Methods for treating glioblastoma with herbal compositions

ABSTRACT

The inventive subject matter relates to methods for treating glioblastoma, comprising administration of a composition comprising therapeutically effective amounts of supercritical extracts of rosemary, turmeric, oregano and ginger; and therapeutically effective amounts of hydroalcoholic extracts of holy basil, ginger, turmeric,  Scutellaria baicalensis , rosemary, green tea, huzhang, Chinese goldthread, and barberry. The inventive subject matter further relates to methods for modulating gene expression of genes selected from the group consisting of interleukin-1α, interleukin-1β, heme oxygenase 1, aldo-keto reductase family 1, member C2, colony stimulating factor 3, leukemia inhibitory factor, heat shock 70 kDa protein, and combinations thereof, by administration of an effective amount of said compositions.

This application claims benefit of U.S. Provisional Application Ser. No.60/609,265 filed on Sep. 14, 2004, the entire contents of which arehereby incorporated.

BACKGROUND OF THE INVENTIVE SUBJECT MATTER

1. Field of the Inventive Subject Matter

The inventive subject matter relates to novel methods for treatingglioblastoma, comprising administration of a composition comprisingtherapeutically effective amounts of supercritical extracts of rosemary,turmeric, oregano and ginger; and therapeutically effective amounts ofhydroalcoholic extracts of holy basil, ginger, turmeric, Scutellariabaicalensis, rosemary, green tea, huzhang, Chinese goldthread, andbarberry. The inventive subject matter further relates to methods formodulating gene expression of genes selected from the group consistingof interleukin-1α, interleukin-1β, heme oxygenase 1, aldo-keto reductasefamily 1, member C2, colony stimulating factor 3, leukemia inhibitoryfactor, heat shock 70 kDa protein, and combinations thereof, byadministration of an effective amount of said compositions.

2. Background

Glioblastoma. Glioblastoma multiforme (GBM) is the most malignant,incurable brain tumor and most patients die in less than a year.Tumorigenesis of GBM is characterized by elevated production ofarachidonic acid-derived lipids called eicosanoids. These moleculesstimulate development of peritumoral brain edema and tumor progression.Glucocorticoids are the most effective drugs that are currently used totreat brain edema, but are associated with numerous side effectsincluding interference with the efficacy of chemotherapy. Because theavailable therapies are not very effective, more than 50% of patientswith GBM use complementary and alternative medicine (CAM) approaches ofwhich herbal therapies are the most commonly used.

5-lipoxygenase Inhibitors. The 5-lipoxygenase (5-LO) pathway is one ofat least four lipoxygenase pathways of arachidonic acid metabolism. The5-lipoxygenase pathway consists of enzymes that regulate a series ofbiochemical reactions that result in the transformation of arachidonicacid to leukotriene A4, which can then be further metabolized toleukotriene B₄ or to leukotriene C₄. Activation of the 5-LO pathwayleads to the biosynthesis of proinflammatory leukotriene lipidmediators, while inhibition of the 5-LO pathway may haveanti-inflammatory effects.

Some compounds which inhibit 5-lipoxygenase have been described in U.S.Pat. Nos. 6,653,311, 6,455,541, 6,399,105, 6,121,323, 5,342,838,5,298,514, 5,145,861, 5,130,483, 4,933,329, and 4,731,382. Drugs such asMK-886(3-(1-(4-chlorobenzyl)-3-tert-butyl-thio-5-isopropylindol-2-yl)-2,2-dimethylpropanoic acid), L-656,224((7-chloro-2-[4-methoxypenyl]methyl)-3-methyl-5-propyl-4-benzofuranol),pentacyclic triterpene acetyl-11-keto-β-boswellic acid, PF-5901,Zileuton, and tepoxalin are intended to selectively inhibit5-lipoxygenase. However, these drugs appear to have long term sideeffects. There is thus a continuing need for 5-lipoxygenase inhibitorswhich avoid side effects associated with current compositions.

Cyclooxygenase Inhibitors. Cyclooxygenase is an enzyme-protein complexwith a variety of biochemical actions. There are at least three primaryCOX isoenzymes, COX-1, COX-2, and COX-3. COX-1 is a constitutive enzyme,produced at a reasonably consistent level at all times. It plays animportant role in, for example, gastrointestinal protection, kidneyfunction, and the aggregation of blood platelets. COX-2 production isnot constant; it varies depending on signals from various biochemicalcatalysts. For example, in the case of arthritis inflammation and pain,COX-2 responds to tissue damage by oxidizing arachidonic acid, creatingprostaglandins which in turn produce local inflammation. COX-3 has beenidentified relatively recently (Chandrasekharan, et al., PNAS U.S.A.,99(21):13926-31 (2002)). In humans, COX-3 mRNA is expressed mostabundantly in the cerebral cortex and heart tissues. Cox-3 activity isselectively inhibited by analgesic/antipyretic drugs. It has beensuggested that inhibition of COX-3 could represent a mechanism by whichthese drugs decrease pain and possibly fever.

Prostaglandins play a major role in the inflammatory process and theinhibition of prostaglandin production, especially production of PGG₂,PGH₂, and PGE₂, has been a common target of anti-inflammatory drugdiscovery. However, common non-steroidal anti-inflammatory drugs(hereinafter “NSAIDs”) that are active in reducing theprostaglandin-induced pain and swelling associated with the inflammationprocess are also active in affecting other prostaglandin-regulatedprocesses not associated with the inflammation process.

NSAIDs have been found to prevent the production of prostaglandins byinhibiting enzymes in the human arachidonic acid/prostaglandin pathway,including the cyclooxygenase enzymes. Traditional non-steroidalanti-inflammatory drugs, such as aspirin, work by inhibiting both COX-1and COX-2. Thus, non-specific NSAIDs can have a damaging effect on thegastrointestinal tract, kidneys, and liver; blocking COX-1 can make thestomach lining more vulnerable, and reduced thromboxane production thinsthe blood, making gastrointestinal hemorrhage more likely, and may causeinadequate regulation of cellular immune functions and the secretion ofvarious cytokines. The use of high doses of most common NSAID's canproduce severe side effects, including life threatening ulcers, thatlimit their therapeutic potential.

COX-2 is associated with inflammation and provides a viable target ofinhibition which more effectively reduces inflammation and producesfewer and less drastic side effects. Thus, researchers have beenmotivated to develop selective COX-2 inhibitors to reduce inflammationand relieve pain without the gastrointestinal damage brought on byinhibiting COX-1. In addition, the current scientific understanding inthe art suggests that COX-2 inhibition may serve an important functionin promoting normal cell growth in the colon, pancreas, breast tissue,and other organ systems.

Some compounds which selectively inhibit cyclooxygenase-2 have beendescribed in U.S. Pat. Nos. 5,380,738, 5,344,991, 5,393,790, 5,434,178,5,474,995, 5,510,368 and WO documents WO96/06840, WO96/03388,WO96/03387, WO96/25405, WO95/15316, WO94/15932, WO94/27980, WO95/00501,WO94/13635, WO94/20480, and WO94/26731.

Drugs such as valdecoxib, celecoxib, and rofecoxib are intended toselectively inhibit COX-2 with minimal effect on COX-1. However, despitethe emphasis on COX-2 inhibition, even these drugs appear to haveserious long term side effects, such as the breakdown in digestiveprotective mucus and prevention of normal healing processes. There isthus a continuing need for more specific and non-specific COX-2inhibitors which avoid side effects associated with COX-1 inhibition.

Natural COX-2 Inhibitors. Several herbs have been found to inhibit theCOX-2 enzyme. For example, holy basil has been found to possesssignificant anti-inflammatory properties and is capable of blocking boththe cyclooxygenase and lipoxygenase pathways of arachidonate metabolism.Ursolic acid and oleanolic acid, two of the recognized phytonutrients ofholy basil, have been found to have a significant COX-2 inhibitoryeffect.

Similarly, shogaols and gingerols, pungent components of ginger, havebeen found to inhibit cyclooxygenase. Eugenol, another activeconstituent of several medical herbs, has also been found to be a5-lipoxygenase inhibitor and to possess potent anti-inflammatory and/oranti-rheumatic properties.

Scutellaria baicalensis also has been found to inhibit the COX-2 enzyme.According to the USDA database, green tea contains six constituentshaving cyclooxygenase-inhibitor activity. According to the Napralertdatabase, green tea contains fifty one constituents havinganti-inflammatory activity. The polyphenols in green tea were found tocause a marked reduction in COX-2. Flavan-3-ol derivatives (+)-catechin,also present in green tea, have been reported to be COX-1 and COX-2inhibitors. In addition, salicylic acid, another constituent of greentea, also has been found to be a COX-2 inhibitor.

Berberine, found in barberry and Chinese goldthread, has also been foundto inhibit COX-2 without inhibiting COX-1 activity.

In U.S. Pat. No. 6,387,416, Applicants disclosed the inventivecompositions and their use for reducing inflammation. The contents ofU.S. Pat. No. 6,387,416 are hereby incorporated by reference in theirentirety. Surprisingly, as discussed in greater detail below, it hasbeen determined that the inventive compositions are useful for treatingglioblastoma as well.

Use of COX-2 Inhibitors for Treating Cancer. It has been postulated thatCOX-2 inhibitors may be useful for treating cancer. Yet only a very fewpatents actually disclose the use of COX-2 inhibitors for treating anycancers. In U.S. Pat. No. 5,466,823 to Talley, et al.,(Pyrazol-1-yl)benzene sulfonamides are disclosed as inhibitors ofcyclooxygenase-2, and for use in the treatment of inflammation,arthritis, and pain, and as being useful for preventing colon cancer.However, their use for actually treating colon cancer or for treating orpreventing other neoplasias is not disclosed.

U.S. Pat. No. 6,469,040 to Seibert, et al., discloses a method of usinga specific, disclosed class of cyclooxygenase-2 inhibitor derivatives inpreventing and treating epithelial cell neoplasia in a subject.

U.S. Pat. No. 6,534,540 to Kindness, et al., discloses a combination ofthe proprietary HMG-CoA reductase inhibitor lovastatin and theproprietary COX-2 inhibitor rofecoxib for the treatment of cancer,especially glioblastoma, and a method of treatment of cancer, especiallyglioblastoma, by that combination.

Based on the limited body of art disclosing the use of COX-2 inhibitorsfor treating any cancer, and the need for effective treatments forglioblastoma in particular, it is apparent that there is a great andimmediate need for new COX-2 inhibitors for treating glioblastoma. Thisneed is met by the inventive methods and compositions, which treatglioblastoma without significant side effects.

SUMMARY OF THE INVENTIVE SUBJECT MATTER

The present inventive subject matter relates to a method for treatingglioblastoma in a subject, comprising the step of administering aneffective amount of a composition to said subject to treat or preventsaid glioblastoma, said composition comprising therapeutically effectiveamounts of supercritical extracts of rosemary, turmeric, oregano andginger; and therapeutically effective amounts of hydroalcoholic extractsof holy basil, ginger, turmeric, Scutellaria baicalensis, rosemary,green tea, huzhang, Chinese goldthread, and barberry.

The present inventive subject matter further relates to a method fortreating glioblastoma in a subject, comprising the step of administeringan effective amount of a composition to said subject to treat saidtumor, said composition comprising therapeutically effective amounts ofsupercritical extracts of rosemary, turmeric, oregano and ginger; andtherapeutically effective amounts of hydroalcoholic extracts of holybasil, ginger, turmeric, Scutellaria baicalensis, rosemary, green tea,huzhang, Chinese goldthread, and barberry.

In addition, the present inventive subject matter is drawn to a methodfor treating side effects associated with glioblastoma in a subject,comprising the step of administering an effective amount of acomposition to said subject to treat said side effects, said compositioncomprising therapeutically effective amounts of supercritical extractsof rosemary, turmeric, oregano and ginger; and

therapeutically effective amounts of hydroalcoholic extracts of holybasil, ginger, turmeric, Scutellaria baicalensis, rosemary, green tea,huzhang, Chinese goldthread, and barberry.

Further, the present inventive subject matter relates to methods formodulating gene expression in a target cell of one or more genesselected from the group consisting of: interleukin-1α, interleukin-1β,heme oxygenase 1, aldo-keto reductase family 1, member C2, colonystimulating factor 3, leukemia inhibitory factor, heat shock 70 kDaprotein, and combinations thereof, comprising administration of aneffective amount of a composition comprising therapeutically effectiveamounts of supercritical extracts of rosemary, turmeric, oregano andginger; and therapeutically effective amounts of hydroalcoholic extractsof holy basil, ginger, turmeric, Scutellaria baicalensis, rosemary,green tea, huzhang, Chinese goldthread, and barberry.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a photograph which depicts induction of heme oxygenase-1protein expression in U87 glioblastoma cells produced by the inventivecompositions, as described in Example 6.

FIG. 2 is a series of three graphs which depict control and experimentalU87 GBM cell apoptosis, as described in Example 2.

DETAILED DESCRIPTION OF THE INVENTIVE SUBJECT MATTER Definitions

The term “therapeutically effective amount” as used herein refers tothat amount of the extract which will contribute to the cancer-treatingability of the composition.

The term “treating” as used herein refers to partial or total inhibitionof the growth, spreading, or metastasis of glioblastoma, as well aspartial or total destruction of the cancer cells. The term “treating”includes the reduction or elimination of glioblastoma, and also thereduction in the incidence of the disease.

The term “preventing” as used herein refers to either preventing theonset of glioblastoma, or preventing the onset of a preclinicallyevident stage of glioblastoma in individuals at risk. Also intended tobe encompassed by this definition is the prevention of initiation formalignant cells, and the arrest or reversal of the progression ofpremalignant cells to malignant cells. “Preventing” also includes theprevention of growth or spreading of the glioblastoma. This includesprophylactic treatment of those at risk of developing a glioblastoma.

The term “supercritical gas” or “supercritical fluid” as used hereinrefers to a gas is that heated to a temperature critical point, overwhich the gas will maintain its gaseous state and not turn to a liquidregardless of pressure. A gas heated to a temperature above its criticalpoint will become very dense on compression, so that its characteristicsresemble those of a fluid, but will not become liquid. Carbon dioxide iscommonly used in applications requiring a supercritical fluid. Thegeneral properties of supercritical fluids and the general use ofsupercritical fluids in extraction processes are described in, e.g.Taylor, Supercritical Fluid Extraction, Wiley, 1996; McHugh andKrukonis, Supercritical Fluid Extraction: Principles and Practice, 2nded., Butterworth-Heinemann, 1994; and Williams and Clifford,Supercritical Fluid Methods and Protocols, Humana Press, 2000, thecontents of which are incorporated by reference herein.

The term “supercritical extraction” as used herein refers to thetechnique in which hydrophobic compounds can be extracted from samplesutilizing a supercritical fluid. The solvation power of a supercriticalfluid is increased as the pressure and temperature are increased abovetheir critical points, producing an effective solvent for the isolationof hydrophobic molecules.

The term “hydroalcoholic extraction” as used herein refers to thetechnique in which hydrophilic compounds can be extracted from a sampleutilizing a solution of alcohol and water, followed by evaporation ofthe solution to produce a extract consisting of dissolved solids.

The term “neoplasia” as used herein refers broadly to neoplastic,pre-malignant, and proliferative disease, including specifically benign,premalignant, or malignant neoplasms in individuals with or without anyprior history or diagnosis of neoplastic, pre-malignant, orproliferative disease. The term “neoplasia” includes neoplasia thatproduce prostaglandins or express a cyclooxygenase, including bothbenign and cancerous tumors, growths, and polyps.

The term “glioblastoma” as used herein refers broadly to neuroglia celltumors and other malignant tumors or neoplasia of the central nervoussystem.

The term “subject” as used herein refers to any human or mammal subjectwho has a glioblastoma, preferably a human subject. For methods ofprevention, the subject is any human or animal subject, preferably ahuman subject, who is at risk for developing glioblastoma. The subjectmay be at risk due to exposure to carcinogenic agents, being geneticallypredisposed to have a glioblastoma, and the like.

The term “cyclooxygenase-2 inhibitor” or “COX-2 inhibitor” as usedherein refers to a compound or composition which is able to inhibitcyclooxygenase-2 without adverse inhibition of cyclooxygenase-1.

Methods for Treating Glioblastoma

Diet is considered as one of the most important modifiable factors incoronary heart disease and carcinogenesis. Recent evidence indicatesthat modulation of inflammation by compounds found in many herbs andspices may be one of the mechanisms by which diet influences developmentand progression of these common chronic conditions.

Both arachidonic acid and its precursor, linoleic acid, are present insignificant quantities in animal fats and a variety of vegetable oils.Physiologically, these fatty acids are integral components of cellularmembranes and also function as substrates for the production of animportant group of potent, signaling lipids, termed eicosanoids.Eicosanoids are known to be involved in the initiation of theinflammatory response, fever production, regulation of blood pressure,blood clotting, control of reproductive processes and tissue growth, andregulation of the sleep/wake cycle. Additionally, these powerfulmediators and the enzymes that produce them, cycloxygenases (COX) andlipoxygenases (LO), are implicated in tumor development, progression,and metastasis.

The three main isoforms of cycloxygenase are COX-1, COX-2, and COX-3,and these enzymes are responsible for the production of the group ofeicosanoids, prostaglandins. The COX-1 isoform has many importanthousekeeping functions in the cell, and is therefore constitutivelyproduced throughout the body. COX-2, however, is usually absent untilinduced by specific stimuli. It is therefore not surprising that COX-2is implicated in the progression of many disease states, includingcancer. COX-2 has been found to be present in elevated levels in avariety of cancers, including lung, colon, pancreatic, head and neck,and glioblastoma. As discussed above, COX-3 has only been relativelyrecently identified.

A wide spectrum of human malignancies aberrantly overexpresspro-inflammatory cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LO)enzymes and inhibitors of these molecules may be useful in cancerchemoprevention and treatment. For this application, Applicants examinedthe effects of Zyflamend® herbal preparation (New Chapter, Inc.,Brattleboro, Vt.; hereinafter “the inventive compositions”) on apoptosisand global gene expression of an established human glioblastoma cellline U87.

The centuries old natural remedy of using white willow bark (Salix alba)to provide some pain relief led to the discovery of aspirin, andeventually, to the elucidation of its mechanism of action as a COXinhibitor. Based on this lead and other traditional Eastern medicinalpractices, many researchers have looked to a variety of natural plantextracts and natural products for the discovery of both non-specific COXand specific COX-2 inhibitors. Some herbal extracts and natural productsthat have peaked interest amongst researchers include curcumin, ginger,holy basil, resveratrol, thundergod vine, and berberine from barberryand Chinese goldthread.

Applicants have developed a mixture comprised of herbal extracts, andthe mixture has COX-2 inhibitory activity. Applicants' compositions areunique, in that they are prepared via a supercritical CO₂ extractionprocess. Unlike traditional solvent based extraction methods,supercritical CO₂ extraction allows the natural products in the herbs tobe obtained without leaving chemical residues behind in the preparation.

Surprisingly, in addition to the anti-inflammatory action disclosed inU.S. Pat. No. 6,387,416, Applicants have found that using the inventivecompositions and methods produce apoptosis in glioblastoma cell lines.Applicants also expect that the inventive methods inhibit cell growth incancer cells which have deactivated apoptotic pathways.

The inventive subject matter is based on the discovery that acombination of certain herbs properly extracted and blended inappropriate proportions can used in treating glioblastoma. Thus,Applicants expect that compositions comprising a therapeuticallyeffective amount of extracts of one or more of rosemary, turmeric,oregano, ginger, holy basil, ginger, turmeric, Scutellaria baicalensis,rosemary, green tea, huzhang, Chinese goldthread, or barberry, andcombinations thereof, are effective in treating glioblastoma. Inparticular, the present inventive subject matter relates to a method fortreating glioblastoma in a subject, comprising the step of administeringan effective amount of a preferred composition to said subject to treator prevent said glioblastoma, said composition comprisingtherapeutically effective amounts of supercritical extracts of rosemary,turmeric, oregano and/or ginger; and/or therapeutically effectiveamounts of hydroalcoholic extracts of holy basil, ginger, turmeric,Scutellaria baicalensis, rosemary, green tea, huzhang, Chinesegoldthread, and/or barberry.

In one aspect, said composition is administered orally.

In another preferred embodiment, the orally administered composition isin the form of one or more capsules, one or more tablets, or one or morepills.

In another aspect, the composition comprises:

-   -   (A) from about 4.5% to about 7.5%, and more preferably from        about 5.5% to about 6.5%, by weight of the hydroalcoholic        extract of ginger;    -   (B) from about 5.5% to about 8.5%, and more preferably from        about 6% to about 8%, by weight of the supercritical extract of        ginger;    -   (C) from about 1.0% to about 1.5%, and more preferably from        about 1.2% to about 1.4%, by weight of the supercritical extract        of turmeric;    -   (D) from about 10.0% to about 16.0%, and more preferably from        about 11.5% to about 14.5%, by weight of the supercritical        extract of rosemary;    -   (E) from about 4.0% to about 6.0%, and more preferably from        about 4.5% to about 5.5%, by weight of the supercritical extract        of oregano;    -   (F) from about 10.0% to about 16.0%, and more preferably from        about 11.5% to about 14.5%, by weight of the hydroalcoholic        extract of turmeric;    -   (G) from about 5.5% to about 8.0%, and more preferably from        about 6.0% to about 7.0%, by weight of the hydroalcoholic        extract of rosemary;    -   (H) from about 10.0% to about 16.0%, and more preferably from        about 11.5% to about 14.5%, by weight of the hydroalcoholic        extract of holy basil;    -   (I) from about 10.0% to about 16.0%, and more preferably from        about 11.5% to about 14.5%, by weight of the hydroalcoholic        extract of green tea;    -   (J) from about 8.0% to about 12.0%, and more preferably from        about 9.0% to about 11.0%, by weight of the hydroalcoholic        extract of huzhang;    -   (K) from about 4.0% to about 6.0%, and more preferably from        about 4.5% to about 5.5%, by weight of the hydroalcoholic        extract of Chinese goldthread;    -   (L) from about 4.0% to about 6.0%, and more preferably from        about 4.5% to about 5.5%, by weight of the hydroalcoholic        extract of barberry; and

(M) from about 2.0% to about 3.0%, and more preferably from about 2.25%to about 2.75%, by weight of the hydroalcoholic extract of Scutellariabaicalensis.

The hydroalcoholic extract of ginger used in the present invention ispreferably prepared as follows. The ginger rhizome, which is preferablycryogenically ground to preserve heat sensitive components, is subjectedto supercritical extraction, preferably with carbon dioxide, to obtain:(i) an oil extract, referred to herein as “the supercritical extract” ofginger, containing delicate lipophilic components, and (ii) an oil-freeresidue. The oil-free residue is then extracted in a water/alcohol,preferably water/ethanol, mixture composed of 60-80 parts alcohol and40-20 parts water. The alcohol/water liquid is then evaporated off,leaving a powdered extract residue, referred to herein as “thehydroalcoholic extract” of ginger.

In a preferred aspect, the weight ratio of the supercritical extract ofginger to the hydroalcoholic extract of ginger is from about 0.9:1 toabout 1.4:1.

The supercritical extracts of ginger, rosemary, turmeric and oreganoused in the present invention can be prepared according to knownsupercritical extraction methods, such as disclosed, e.g., in E. Stahl,K. W. Quirin, D. Gerard, Dense Gases for Extraction and Refining,Springer Verlag 4 1988, which is hereby incorporated by referenceherein.

The hydroalcoholic extracts of rosemary, turmeric, holy basil, greentea, huzhang, Chinese goldthread, barberry and Scutellaria baicalensisused in the present invention can be prepared according to conventionalhydroalcoholic extraction techniques. For example, the hydroalcoholicextracts can be prepared by extracting the plant portion in awater/alcohol, preferably water/ethanol, mixture preferably composed of60-80 parts alcohol and 40-20 parts water, and then evaporating off thewater/alcohol liquid, leaving a powdered extract residue referred toherein as “the hydroalcoholic extract”.

In yet another aspect, the weight ratio of the hydroalcoholic extract ofturmeric to the supercritical extract of turmeric is from about 8:1 toabout 12:1.

In an alternate aspect, the weight ratio of the supercritical extract ofrosemary to the hydroalcoholic extract of rosemary is from about 1.6:1to about 2.4:1.

In a still further aspect, the hydroalcoholic extract of gingercomprises from about 2.4% to about 3.6%, more preferably from about 2.7%to about 3.3%, and most preferably about 3.0%, by weight of pungentcompounds.

In another aspect, the supercritical extract of ginger comprises fromabout 24% to about 36%, more preferably from about 27% to about 33%, andmost preferably about 30%, by weight of pungent compounds; and fromabout 6.4% to about 9.6%, more preferably from about 7.2% to about 8.8%,and most preferably about 8%, by weight of zingiberene.

In a further aspect, the supercritical extract of turmeric comprisesfrom about 36% to about 54%, more preferably from about 40.5% to about49.5%, and most preferably about 45%, by weight of turmerones.

In another aspect, the supercritical extract of rosemary comprises fromabout 18.4% to about 27.6%, more preferably from about 20.7% to about25.3%, and most preferably about 23%, by weight of total phenolicantioxidants.

In yet another aspect, the supercritical extract of oregano comprisesfrom about 0.64% to about 0.96%, more preferably from about 0.72% toabout 0.88%, and most preferably about 0.8%, by weight of total phenolicantioxidants.

In a still further aspect, the hydroalcoholic extract of turmericcomprises from about 5.6% to about 8.4%, more preferably from about 6.3%to about 7.7%, and most preferably about 7%, by weight of curcumin.

In another aspect, the hydroalcoholic extract of rosemary comprises fromabout 18.4% to about 27.6%, more preferably from about 20.7% to about25.3%, and most preferably about 23%, by weight of total phenolicantioxidants.

In a further embodiment, the hydroalcoholic extract of holy basilcomprises from about 1.6% to about 2.4%, more preferably from about 1.8%to about 2.2%, and most preferably about 2%, by weight of ursolic acid.

In a further aspect, the hydroalcoholic extract of green tea comprisesfrom about 36% to about 54%, more preferably from about 40.5% to about49.5%, and most preferably about 45%, by weight of polyphenols.

In another aspect, the hydroalcoholic extract of huzhang comprises fromabout 6.4% to about 9.6%, more preferably from about 7.2% to about 8.8%,and most preferably about 8%, by weight of resveratrol.

In another embodiment, the hydroalcoholic extract of Chinese goldthreadcomprises from about 4.8% to about 7.2%, more preferably from about 5.4%to about 6.6%, and most preferably about 6%, by weight of berberine.

In a further aspect, the hydroalcoholic extract of barberry comprisesfrom about 4.8% to about 7.2%, more preferably from about 5.4% to about6.6%, and most preferably about 6%, by weight of berberine.

In an alternate aspect, said composition comprises:

-   -   (A) from about 4.5% to about 7.5% by weight of the        hydroalcoholic extract of ginger, wherein the extract comprises        from about 2.4% to about 3.6% by weight of pungent compounds;    -   (B) from about 5.5% to about 8.5% by weight of the supercritical        extract of ginger, wherein the extract comprises from about 24%        to about 36% by weight of pungent compounds and from about 6.4%        to about 9.6% by weight of zingiberene;    -   (C) from about 1.0% to about 1.5% by weight of the supercritical        extract of turmeric, wherein the extract comprises from about        36% to about 54% by weight of turmerones;    -   (D) from about 10.0% to about 16.0% by weight of the        supercritical extract of rosemary, wherein the extract comprises        from about 18.4% to about 27.6% by weight of total phenolic        antioxidants;    -   (E) from about 4.0% to about 6.0% by weight of the supercritical        extract of oregano, wherein the extract comprises from about        0.64% to about 0.96% by weight of total phenolic antioxidants;    -   (F) from about 10.0% to about 16.0% by weight of the        hydroalcoholic extract of turmeric, wherein the extract        comprises from about 5.6% to about 8.4% by weight of curcumin;    -   (G) from about 5.5% to about 8.0% by weight of the        hydroalcoholic extract of rosemary, wherein the extract        comprises from about 18.4% to about 27.6% by weight of total        phenolic antioxidants;    -   (H) from about 10.0% to about 16.0% by weight of the        hydroalcoholic extract of holy basil, wherein the extract        comprises from about 1.6% to about 2.4% by weight of ursolic        acid;    -   (I) from about 10.0% to about 16.0% by weight of the        hydroalcoholic extract of green tea, wherein the extract        comprises from about 36% to about 54% by weight of polyphenols;    -   (J) from about 8.0% to about 12.0% by weight of the        hydroalcoholic extract of huzhang, wherein the extract comprises        from about 6.4% to about 9.6% by weight of resveratrol;    -   (K) from about 4.0% to about 6.0% by weight of the        hydroalcoholic extract of Chinese goldthread, wherein the        extract from about 4.8% to about 7.2% by weight of berberine;    -   (L) from about 4.0% to about 6.0% by weight of the        hydroalcoholic extract of barberry, wherein the extract from        about 4.8% to about 7.2% by weight of berberine; and    -   (M) from about 2.0% to about 3.0% by weight of the        hydroalcoholic extract of Scutellaria baicalensis;    -   and wherein said composition further comprises:    -   (i) the supercritical extract of ginger and the        post-supercritical hydroalcoholic extract of ginger at a weight        ratio of from about 0.9 to about 1.4 parts of supercritical        extract per 1 part of post-supercritical hydroalcoholic extract;    -   (ii) the hydroalcoholic extract of turmeric and the        supercritical extract of turmeric at a weight ratio of from        about 8 to about 12 parts of hydroalcoholic extract per 1 part        of supercritical extract; and    -   (iii) the supercritical extract of rosemary and the        hydroalcoholic extract of rosemary at a weight ratio of from        about 1.6 to about 2.4 parts of supercritical extract per 1 part        of hydroalcoholic extract.

In a preferred embodiment, the composition is administered in a dailydosage of at least about 700 mg.

In another aspect, the composition is administered on a daily basis forat least 4 weeks.

A still further aspect of the present inventive subject matter is drawnto a method for treating at least one cancerous tumor of the centralnervous system in a subject, comprising the step of administering aneffective amount of a composition to said subject to treat said tumor,said composition comprising therapeutically effective amounts ofsupercritical extracts of rosemary, turmeric, oregano and ginger; andtherapeutically effective amounts of hydroalcoholic extracts of holybasil, ginger, turmeric, Scutellaria baicalensis, rosemary, green tea,huzhang, Chinese goldthread, and barberry.

In a still further embodiment of the present inventive subject matter,the at least one cancerous tumor is detected during surgery on thecentral nervous system of said subject, having not been felt by aphysician on physical examination of said subject.

Yet still further, the present inventive subject matter includes the atleast one cancerous tumor being confined to the central nervous systemof said subject and is detected by a physician on physical examinationof said subject.

An additional aspect of the present invention includes the cancerrelated to said at least one cancerous tumor extends beyond the centralnervous system of said subject, but has not spread to lymph nodes insaid subject.

A further additional aspect of the present inventive subject matter isdirected to the cancer related to said at least one cancerous tumor ismetastasized to regional lymph nodes or other parts of said subject.

A preferred aspect of the present invention is directed to a method fortreating side effects associated with glioblastoma in a subject,comprising the step of administering an effective amount of acomposition to said subject to treat said side effects, said compositioncomprising therapeutically effective amounts of supercritical extractsof rosemary, turmeric, oregano and ginger; and therapeutically effectiveamounts of hydroalcoholic extracts of holy basil, ginger, turmeric,Scutellaria baicalensis, rosemary, green tea, huzhang, Chinesegoldthread, and barberry.

In an alternate aspect, the composition comprises an additional agentselected from the group consisting of antineoplastic agents, growthinhibiting agents, and nutrients.

There are large numbers of antineoplastic agents available in commercialuse, in clinical evaluation and in pre-clinical development, whichoptionally are selected for treatment of glioblastoma by combinationdrug chemotherapy. Such antineoplastic agents fall into several majorcategories: antimetabolite agents, antibiotic-type agents, alkylatingagents, hormonal agents, immunological agents, interferon-type agents,metallomatrix proteases, superoxide dismutase mimics or α_(v)β₃inhibitors. Thus, in a preferred embodiment, said antineoplastic agentis selected from the group consisting of antimetabolite agents,antibiotic-type agents, alkylating agents, hormonal agents,immunological agents, interferon-type agents, metallomatrix proteases,superoxide dismutase mimics, and α_(v)β₃ inhibitors.

One class of antineoplastic agents which may be used in combination withan inventive composition consists of antimetabolite-type antineoplasticagents. Suitable antimetabolite antineoplastic agents may be selectedfrom the group consisting of 5-FU-fibrinogen, acanthifolic acid,aminothiadiazole, brequinar sodium, carmofur, Ciba-Geigy CGP-30694,cyclopentyl cytosine, cytarabine phosphate stearate, cytarabineconjugates, Lilly DATHF, Merrel Dow DDFC, dezaguanine, dideoxycytidine,dideoxyguanosine, didox, Yoshitomi DMDC, doxifluridine, Wellcome EHNA,Merck & Co. EX-015, fazarabine, floxuridine, fludarabine phosphate,5-fluorouracil, N-(2′-furanidyl)-5-fluorouracil, Daiichi Seiyaku FO-152,isopropyl pyrrolizine, Lilly LY-188011, Lilly LY-264618, methobenzaprim,methotrexate, Wellcome MZPES, norspermidine, NCI NSC-127716, NCINSC-264880, NCI NSC-39661, NCI NSC-612567, Warner-Lambert PALA,pentostatin, piritrexim, plicamycin, Asahi Chemical PL-AC, TakedaTAC-788, thioguanine, tiazofurin, Erbamont TIF, trimetrexate, tyrosinekinase inhibitors, tyrosine protein kinase inhibitors, Taiho UFT, anduricytin.

A second class of antineoplastic agents which may be used in combinationwith an inventive composition consists of alkylating-type antineoplasticagents. Suitable alkylating-type antineoplastic agents may be selectedfrom the group consisting of Shionogi 254-S, aldo-phosphamide analogues,altretamine, anaxirone, Boehringer Mannheim BBR-2207, bestrabucil,budotitane, Wakunaga CA-102, carboplatin, carmustine, Chinoin-139,Chinoin-153, chlorambucil, cisplatin, cyclophosphamide, AmericanCyanamid CL-286558, Sanofi CY-233, cyplatate, Degussa D-19-384, SumimotoDACHP(Myr)2, diphenylspiromustine, diplatinum cytostatic, Erbadistamycin derivatives, Chugai DWA-2114R, ITI E09, elmustine, ErbamontFCE-24517, estramustine phosphate sodium, fotemustine, Unimed G-6-M,Chinoin GYKI-17230, hepsul-fam, ifosfamide, iproplatin, lomustine,mafosfamide, mitolactol, Nippon Kayaku NK-121, NCI NSC-264395, NCINSC-342215, oxaliplatin, Upjohn PCNU, prednimustine, Proter PTT-119,ranimustine, semustine, SmithKline SK&F-101772, Yakult Honsha SN-22,spiromustine, Tanabe Seiyaku TA-077, tauromustine, temozolomide,teroxirone, tetraplatin, and trimelamol.

A third class of antineoplastic agents which may be used in combinationwith an inventive composition consists of antibiotic-type antineoplasticagents. Suitable antibiotic-type antineoplastic agents may be selectedfrom the group consisting of Taiho 4181-A, aclarubicin, actinomycin D,actinoplanone, Erbamont ADR-456, aeroplysinin derivative, AjinomotoAN-201-II, Ajinomoto AN-3, Nippon Soda anisomycins, anthracycline,azino-mycin-A, bisucaberin, Bristol-Myers BL-6859, Bristol-MyersBMY-25067, Bristol-Myers BMY-25551, Bristol-Myers BMY-26605,Bristol-Myers BMY-27557, Bristol-Myers BMY-28438, bleomycin sulfate,bryostatin-1, Taiho C-1027, calichemycin, chromoximycin, dactinomycin,daunorubicin, Kyowa Hakko DC-102, Kyowa Hakko DC-79, Kyowa Hakko DC-88A,Kyowa Hakko DC89-A1, Kyowa Hakko DC92-B, ditrisarubicin B, ShionogiDOB-41, doxorubicin, doxorubicin-fibrinogen, elsamicin-A, epirubicin,erbstatin, esorubicin, esperamicin-A1, esperamicin-Alb, ErbamontFCE-21954, Fujisawa FK-973, fostriecin, Fujisawa FR-900482, glidobactin,gregatin-A, grincamycin, herbimycin, idarubicin, illudins, kazusamycin,kesarirhodins, Kyowa Hakko KM-5539, Kirin Brewery KRN-8602, Kyowa HakkoKT-5432, Kyowa Hakko KT-5594, Kyowa Hakko KT-6149, American CyanamidLL-D49194, Meiji Seika ME 2303, menogaril, mitomycin, mitoxantrone,SmithKline M-TAG, neoenactin, Nippon Kayaku NK-313, Nippon KayakuNKT-01, SRI International NSC-357704, oxalysine, oxaunomycin,peplomycin, pilatin, pirarubicin, porothramycin, pyrindamycin A, TobishiRA-I, rapamycin, rhizoxin, rodorubicin, sibanomicin, siwenmycin,Sumitomo SM-5887, Snow Brand SN-706, Snow Brand SN-07, sorangicin-A,sparsomycin, SS Pharmaceutical SS-21020, SS Pharmaceutical SS-7313B, SSPharmaceutical SS-9816B, steffimycin B, Taiho 4181-2, talisomycin,Takeda TAN-868A, terpentecin, thrazine, tricrozarin A, Upjohn U-73975,Kyowa Hakko UCN-10028A, Fujisawa WF-3405, Yoshitomi Y-25024, andzorubicin.

A fourth class of antineoplastic agents which may be used in combinationwith an inventive composition consists of a miscellaneous family ofantineoplastic agents selected from the group consisting ofalpha-carotene, alpha-difluoromethyl-arginine, acitretin, Biotec AD-5,Kyorin AHC-52, alstonine, amonafide, amphethinile, amsacrine, Angiostat,ankinomycin, anti-neoplaston A10, antineoplaston A2, antineoplaston A3,antineoplaston A5, antineoplaston AS2-1, Henkel APD, aphidicolinglycinate, asparaginase, Avarol, baccharin, batracylin, benfluron,benzotript, Ipsen-Beaufour BIM-23015, bisantrene, Bristo-MyersBMY-40481, Vestar boron-10, bromofosfamide, Wellcome BW-502, WellcomeBW-773, caracemide, carmethizole hydrochloride, Ajinomoto CDAF,chlorsulfaquinoxalone, Chemes CHX-2053, Chemex CHX-100, Warner-LambertCI-921, Warner-Lambert CI-937, Warner-Lambert CI-941, Warner-LambertCI-958, clanfenur, claviridenone, ICN compound 1259, ICN compound 4711,Contracan, Yakult Honsha CPT-11, crisnatol, curaderm, cytochalasin B,cytarabine, cytocytin, Merz D-609, DABIS maleate, dacarbazine,datelliptinium, didemnin-B, dihaematoporphyrin ether, dihydrolenperone,dinaline, distamycin, Toyo Pharmar DM-341, Toyo Pharmar DM-75, DaiichiSeiyaku DN-9693, elliprabin, elliptinium acetate, Tsumura EPMTC,ergotamine, etoposide, etretinate, fenretinide, Fujisawa FR-57704,gallium nitrate, genkwadaphnin, Chugai GLA-43, Glaxo GR-63178, grifolanNMF-5N, hexadecylphosphocholine, Green Cross HO-221, homoharringtonine,hydroxyurea, BTG ICRF-187, ilmofosine, isoglutamine, isotretinoin,Otsuka JI-36, Ramot K-477, Otsuak K-76COONa, Kureha Chemical K-AM, MECTCorp KI-8110, American Cyanamid L-623, leukoregulin, lonidamine,Lundbeck LU-23-112, Lilly LY-186641, NCI (US) MAP, marycin, Merrel DowMDL-27048, Medco MEDR-340, merbarone, merocyanine derivatives,methylanilinoacridine, Molecular Genetics MGI-136, minactivin,mitonafide, mitoquidone, mopidamol, motretinide, Zenyaku Kogyo MST-16,N-(retinoyl)amino acids, Nisshin Flour Milling N-021,N-acylated-dehydroalanines, nafazatrom, Taisho NCU-190, nocodazolederivative, Normosang, NCI NSC-145813, NCI NSC-361456, NCI NSC-604782,NCI NSC-95580, octreotide, Ono ONO-112, oquizanocine, Akzo Org-10172,pancratistatin, pazelliptine, Warner-Lambert PD-111707, Warner-LambertPD-115934, Warner-Lambert PD-131141, Pierre Fabre PE-1001, ICRT peptideD, piroxantrone, polyhaematoporphyrin, polypreic acid, Efamol porphyrin,probimane, procarbazine, proglumide, Invitron protease nexin I, TobishiRA-700, razoxane, Sapporo Breweries RBS, restrictin-P, retelliptine,retinoic acid, Rhone-Poulenc RP-49532, Rhone-Poulenc RP-56976,SmithKline SK&F-104864, Sumitomo SM-108, Kuraray SMANCS, SeaPharmSP-10094, spatol, spirocyclopropane derivatives, spirogermanium, Unimed,SS Pharmaceutical SS-554, strypoldinone, Stypoldione, Suntory SUN 0237,Suntory SUN 2071, superoxide dismutase, Toyama T-506, Toyama T-680,taxol, Teijin TEI-0303, teniposide, thaliblastine, Eastman Kodak TJB-29,tocotrienol, Topostin, Teijin TT-82, Kyowa Hakko UCN-01, Kyowa HakkoUCN-1028, ukrain, Eastman Kodak USB-006, vinblastine sulfate,vincristine, vindesine, vinestramide, vinorelbine, vintriptol,vinzolidine, withanolides, and Yamanouchi YM-534.

Examples of radioprotective agents which may be used in the combinationchemotherapy of this invention are AD-5, adchnon, amifostine analogues,detox, dimesna, 1-102, MM-159, N-acylated-dehydroalanines,TGF-Genentech, tiprotimod, amifostine, WR-151327, FUT-187, ketoprofentransdermal, nabumetone, superoxide dismutase (Chiron), and superoxidedismutase Enzon.

Thus, in a further preferred embodiment, said antineoplastic agent isselected from the group consisting of 5-FU-fibrinogen, acanthifolicacid, aminothiadiazole, brequinar sodium, carmofur, Ciba-GeigyCGP-30694, cyclopentyl cytosine, cytarabine phosphate stearate,cytarabine conjugates, Lilly DATHF, Merrel Dow DDFC, dezaguanine,dideoxycytidine, dideoxyguanosine, didox, Yoshitomi DMDC, doxifluridine,Wellcome EHNA, Merck & Co. EX-015, fazarabine, floxuridine, fludarabinephosphate, 5-fluorouracil, N-(2′-furanidyl)-5-fluorouracil, DaiichiSeiyaku FO-152, isopropyl pyrrolizine, Lilly LY-188011, Lilly LY-264618,methobenzaprim, methotrexate, Wellcome MZPES, norspermidine, NCINSC-127716, NCI NSC-264880, NCI NSC-39661, NCI NSC-612567,Warner-Lambert PALA, pentostatin, piritrexim, plicamycin, Asahi ChemicalPL-AC, Takeda TAC-788, thioguanine, tiazofurin, Erbamont TIF,trimetrexate, tyrosine kinase inhibitors, tyrosine protein kinaseinhibitors, Taiho UFT, uricytin, Shionogi 254-S, aldo-phosphamideanalogues, altretamine, anaxirone, Boehringer Mannheim BBR-2207,bestrabucil, budotitane, Wakunaga CA-102, carboplatin, carmustine,Chinoin-139, Chinoin-153, chlorambucil, cisplatin, cyclophosphamide,American Cyanamid CL-286558, Sanofi CY-233, cyplatate, Degussa D-19-384,Sumimoto DACHP(Myr)2, diphenylspiromustine, diplatinum cytostatic, Erbadistamycin derivatives, Chugai DWA-2114R, ITI E09, elmustine, ErbamontFCE-24517, estramustine phosphate sodium, fotemustine, Unimed G-6-M,Chinoin GYKI-17230, hepsul-fam, ifosfamide, iproplatin, lomustine,mafosfamide, mitolactol, Nippon Kayaku NK-121, NCI NSC-264395, NCINSC-342215, oxaliplatin, Upjohn PCNU, prednimustine, Proter PTT-119,ranimustine, semustine, SmithKline SK&F-101772, Yakult Honsha SN-22,spiromustine, Tanabe Seiyaku TA-077, tauromustine, temozolomide,teroxirone, tetraplatin, trimelamol, Taiho 4181-A, aclarubicin,actinomycin D, actinoplanone, Erbamont ADR-456, aeroplysinin derivative,Ajinomoto AN-201-II, Ajinomoto AN-3, Nippon Soda anisomycins,anthracycline, azino-mycin-A, bisucaberin, Bristol-Myers BL-6859,Bristol-Myers BMY-25067, Bristol-Myers BMY-25551, Bristol-MyersBMY-26605, Bristol-Myers BMY-27557, Bristol-Myers BMY-28438, bleomycinsulfate, bryostatin-1, Taiho C-1027, calichemycin, chromoximycin,dactinomycin, daunorubicin, Kyowa Hakko DC-102, Kyowa Hakko DC-79, KyowaHakko DC-88A, Kyowa Hakko DC89-A1, Kyowa Hakko DC92-B, ditrisarubicin B,Shionogi DOB-41, doxorubicin, doxorubicin-fibrinogen, elsamicin-A,epirubicin, erbstatin, esorubicin, esperamicin-A1, esperamicin-Alb,Erbamont FCE-21954, Fujisawa FK-973, fostriecin, Fujisawa FR-900482,glidobactin, gregatin-A, grincamycin, herbimycin, idarubicin, illudins,kazusamycin, kesarirhodins, Kyowa Hakko KM-5539, Kirin Brewery KRN-8602,Kyowa Hakko KT-5432, Kyowa Hakko KT-5594, Kyowa Hakko KT-6149, AmericanCyanamid LL-D49194, Meiji Seika ME 2303, menogaril, mitomycin,mitoxantrone, SmithKline M-TAG, neoenactin, Nippon Kayaku NK-313, NipponKayaku NKT-01, SRI International NSC-357704, oxalysine, oxaunomycin,peplomycin, pilatin, pirarubicin, porothramycin, pyrindamycin A, TobishiRA-I, rapamycin, rhizoxin, rodorubicin, sibanomicin, siwenmycin,Sumitomo SM-5887, Snow Brand SN-706, Snow Brand SN-07, sorangicin-A,sparsomycin, SS Pharmaceutical SS-21020, SS Pharmaceutical SS-7313B, SSPharmaceutical SS-9816B, steffimycin B, Taiho 4181-2, talisomycin,Takeda TAN-868A, terpentecin, thrazine, tricrozarin A, Upjohn U-73975,Kyowa Hakko UCN-10028A, Fujisawa WF-3405, Yoshitomi Y-25024, zorubicin,alpha-carotene, alpha-difluoromethyl-arginine, acitretin, Biotec AD-5,Kyorin AHC-52, alstonine, amonafide, amphethinile, amsacrine, Angiostat,ankinomycin, anti-neoplaston A10, antineoplaston A2, antineoplaston A3,antineoplaston A5, antineoplaston AS2-1, Henkel APD, aphidicolinglycinate, asparaginase, Avarol, baccharin, batracylin, benfluron,benzotript, Ipsen-Beaufour BIM-23015, bisantrene, Bristo-MyersBMY-40481, Vestar boron-10, bromofosfamide, Wellcome BW-502, WellcomeBW-773, caracemide, carmethizole hydrochloride, Ajinomoto CDAF,chlorsulfaquinoxalone, Chemes CHX-2053, Chemex CHX-100, Warner-LambertCI-921, Warner-Lambert CI-937, Warner-Lambert CI-941, Warner-LambertCI-958, clanfenur, claviridenone, ICN compound 1259, ICN compound 4711,Contracan, Yakult Honsha CPT-11, crisnatol, curaderm, cytochalasin B,cytarabine, cytocytin, Merz D-609, DABIS maleate, dacarbazine,datelliptinium, didemnin-B, dihaematoporphyrin ether, dihydrolenperone,dinaline, distamycin, Toyo Pharmar DM-341, Toyo Pharmar DM-75, DaiichiSeiyaku DN-9693, elliprabin, elliptinium acetate, Tsumura EPMTC,ergotamine, etoposide, etretinate, fenretinide, Fujisawa FR-57704,gallium nitrate, genkwadaphnin, Chugai GLA-43, Glaxo GR-63178, grifolanNMF-5N, hexadecylphosphocholine, Green Cross HO-221, homoharringtonine,hydroxyurea, BTG ICRF-187, ilmofosine, isoglutamine, isotretinoin,Otsuka JI-36, Ramot K-477, Otsuak K-76COONa, Kureha Chemical K-AM, MECTCorp KI-8110, American Cyanamid L-623, leukoregulin, lonidamine,Lundbeck LU-23-112, Lilly LY-186641, NCI (US) MAP, marycin, Merrel DowMDL-27048, Medco MEDR-340, merbarone, merocyanine derivatives,methylanilinoacridine, Molecular Genetics MGI-136, minactivin,mitonafide, mitoquidone, mopidamol, motretinide, Zenyaku Kogyo MST-16,N-(retinoyl)amino acids, Nisshin Flour Milling N-021,N-acylated-dehydroalanines, nafazatrom, Taisho NCU-190, nocodazolederivative, Normosang, NCI NSC-145813, NCI NSC-361456, NCI NSC-604782,NCI NSC-95580, octreotide, Ono ONO-112, oquizanocine, Akzo Org-10172,pancratistatin, pazelliptine, Warner-Lambert PD-111707, Warner-LambertPD-115934, Warner-Lambert PD-131141, Pierre Fabre PE-1001, ICRT peptideD, piroxantrone, polyhaematoporphyrin, polypreic acid, Efamol porphyrin,probimane, procarbazine, proglumide, Invitron protease nexin I, TobishiRA-700, razoxane, Sapporo Breweries RBS, restrictin-P, retelliptine,retinoic acid, Rhone-Poulenc RP-49532, Rhone-Poulenc RP-56976,SmithKline SK&F-104864, Sumitomo SM-108, Kuraray SMANCS, SeaPharmSP-10094, spatol, spirocyclopropane derivatives, spirogermanium, Unimed,SS Pharmaceutical SS-554, strypoldinone, Stypoldione, Suntory SUN 0237,Suntory SUN 2071, superoxide dismutase, Toyama T-506, Toyama T-680,taxol, Teijin TEI-0303, teniposide, thaliblastine, Eastman Kodak TJB-29,tocotrienol, Topostin, Teijin TT-82, Kyowa Hakko UCN-01, Kyowa HakkoUCN-1028, ukrain, Eastman Kodak USB-006, vinblastine sulfate,vincristine, vindesine, vinestramide, vinorelbine, vintriptol,vinzolidine, withanolides, Yamanouchi YM-534, AD-5, adchnon, amifostineanalogues, detox, dimesna, 1-102, MM-159, N-acylated-dehydroalanines,TGF-Genentech, tiprotimod, amifostine, WR-151327, FUT-187, ketoprofentransdermal, nabumetone, and superoxide dismutase.

A benefit provided by the inventive compositions is the utilization ofsupercritical extraction, an innovative technology for extracting herbsat low temperature without the use of industrial chemical solvents. Suchextraction process allows for the highest potency of active compounds inthe extracts, as much as 250 times the potency of the original freshplant material.

Set forth in Table I is a preferred embodiment of the orallyadministered composition, excluding inactive ingredients, as used in theinventive methods. The amounts recited in Table I represent thepreferred dosage of the ingredients listed.

TABLE I Plant Amount Herb Type Of Extract Part (mg) Rosemarysupercritical leaf 100 Rosemary hydroalcoholic (23% TPA - 34.5 mg) leaf50 Turmeric supercritical (45% turmerones - 4.5 mg) rhizome 10 Turmerichydroalcoholic (7% curcumin - 7 mg) rhizome 100 Ginger supercritical(30% pungent compounds - rhizome 54 16.2 mg 8% zingiberene - 4.3 mg)Ginger hydroalcoholic (3% pungent rhizome 46 compounds - 1.4 mg) Holybasil hydroalcoholic (2% ursolic acid - 2 mg) leaf 100 Green teahydroalcoholic (45% polyphenols - leaf 100 45 mg) Huzhang hydroalcoholic(8% resveratrol - root & 80 6.4 mg) rhizome Chinese hydroalcoholic (6%berberine - 2.4 mg) root 40 Goldthread Barberry hydroalcoholic (6%berberine - 2.4 mg) root 40 Oregano supercritical (0.8% TPA - 0.32 mg)leaf 40 Scutellaria hydroalcoholic (5:1) root 20 Baicalensis

Preferably, the composition set forth in Table I also includes extravirgin olive oil and yellow beeswax.

The inventive methods use a therapeutically effective amount of theactive compositions indicated above. This effective amount willgenerally comprise from about 0.1 mg to about 100 mg of the active agentper kilogram of patient body weight per day. This effective amount canvary depending upon the physical status of the patient and other factorswell known in the art. Moreover, it will be understood that this dosageof active agent can be administered in a single or multiple dosage unitsto provide the desired therapeutic effect. If desired, other therapeuticagents can be employed in conjunction with those provided by the presentinventive subject matter.

The inventive methods use compositions which are preferably delivered tothe patient by means of a pharmaceutically acceptable carrier. Suchcarriers are well known in the art and generally will be in either solidor liquid form. Solid form pharmaceutical preparations which may beprepared according to the present inventive subject matter includepowders, tablets, dispersible granules, capsules, and cachets. Ingeneral, solid form preparations will comprise from about 5% to about90% by weight of the active agent.

A solid carrier can be one or more substances which may also act asdiluents, flavoring agents, solubilizers, lubricants, suspending agents,binders or tablet disintegrating agents; it can also be encapsulatingmaterial. In powders, the carrier is a finely divided solid which is inadmixture with the viscous active compound. In tablets, the activecompound is mixed with a carrier having the necessary binding propertiesin suitable proportions and compacted to the shape and size desired.Suitable solid carriers include magnesium carbonate, magnesium stearate,talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth,methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoabutter, and the like. The term “preparation” is intended to include theformulation of the active compound with encapsulating materials as acarrier which may provide a capsule in which the active component (withor without other carriers) is surrounded by carrier, which is thus inassociation with it. Similarly, cachets are included. Tablets, powders,cachets, and capsules can be used as solid dosage forms suitable fororal administration. If desired for reasons of convenience or patientacceptance, pharmaceutical tablets prepared according to the inventivesubject matter may be provided in chewable form, using techniques wellknown in the art.

Also contemplated as suitable carriers are solid form preparations whichare intended to be converted, shortly before use, to liquid formpreparations for either oral or parenteral administration. Such liquidforms include solutions, suspensions, and emulsions. These particularsolid form preparations are most conveniently provided in unit dose formand as such are used to provide a single liquid dosage unit.Alternately, sufficient solid may be provided so that after conversionto liquid form, multiple individual liquid doses may be obtained bymeasuring predetermined volumes of the liquid form preparation as with asyringe, teaspoon, or other volumetric container. When multiple liquiddoses are so prepared, it is preferred to maintain the unused portion ofsaid liquid doses at low temperature (i.e., under refrigeration) inorder to retard possible decomposition. The solid form preparationsintended to be converted to liquid form may contain, in addition to theactive material, flavorants, colorants, stabilizers, buffers, artificialand natural sweeteners, dispersants, thickeners, solubilizing agents,and the like. The liquid utilized for preparing useful liquid formpreparations may be water, isotonic water, ethanol, glycerine, propyleneglycol, and the like as well as mixtures thereof. Naturally, the liquidutilized will be chosen with regard to the route of administration. Forexample, liquid preparations containing large amounts of ethanol are notsuitable for parenteral use.

The pharmaceutical preparation may also be in a unit dosage form. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, for example, packeted tablets, capsules, and powders invials or ampoules. The unit dosage form can also be a capsule, cachet,or tablet itself or it can be the appropriate number of any of these inpackaged form.

The pharmaceutical preparations of the inventive subject matter mayinclude one or more preservatives well known in the art, such as benzoicacid, sorbic acid, methylparaben, propylparaben and ethylenediaminetetraacetic acid (EDTA). Preservatives are generally present in amounts up toabout 1% and preferably from about 0.05 to about 0.5% by weight of thepharmaceutical composition.

Useful buffers for purposes of the inventive subject matter includecitric acid-sodium citrate, phosphoric acid-sodium phosphate, and aceticacid-sodium acetate in amounts up to about 1% and preferably from about0.05 to about 0.5% by weight of the pharmaceutical composition. Usefulsuspending agents or thickeners include cellulosics likemethylcellulose, carageenans like alginic acid and its derivatives,xanthan gums, gelatin, acacia, and microcrystalline cellulose in amountsup to about 20% and preferably from about 1% to about 15% by weight ofthe pharmaceutical composition.

Sweeteners which may be employed include those sweeteners, both naturaland artificial, well known in the art. Sweetening agents such asmonosaccharides, disaccharides and polysaccharides such as xylose,ribose, glucose, mannose, galactose, fructose, dextrose, sucrose,maltose, partially hydrolyzed starch or corn syrup solids and sugaralcohols such as sorbitol, xylitol, mannitol and mixtures thereof may beutilized in amounts from about 10% to about 60% and preferably fromabout 20% to about 50% by weight of the pharmaceutical composition.Water soluble artificial sweeteners such as saccharin and saccharinsalts such as sodium or calcium, cyclamate salts, acesulfame-K,aspartame and the like and mixtures thereof may be utilized in amountsfrom about 0.001% to about 5% by weight of the composition.

Flavorants which may be employed in the pharmaceutical products of theinventive subject matter include both natural and artificial flavors,and mints such as peppermint, menthol, vanilla, artificial vanilla,chocolate, artificial chocolate, cinnamon, various fruit flavors, bothindividually and mixed, in amounts from about 0.5% to about 5% by weightof the pharmaceutical composition.

Colorants useful in the present inventive subject matter includepigments which may be incorporated in amounts of up to about 6% byweight of the composition. A preferred pigment, titanium dioxide, may beincorporated in amounts up to about 1%. Also, the colorants may includeother dyes suitable for food, drug and cosmetic applications, known asF.D.&C. dyes and the like. Such dyes are generally present in amounts upto about 0.25% and preferably from about 0.05% to about 0.2% by weightof the pharmaceutical composition. A full recitation of all F.D.&C. andD.&C. dyes and their corresponding chemical structures may be found inthe Kirk-Othmer Encyclopedia of Chemical Technology, in Volume 5, atpages 857-884, which text is accordingly incorporated herein byreference.

Useful solubilizers include alcohol, propylene glycol, polyethyleneglycol and the like and may be used to solubilize the flavors.Solubilizing agents are generally present in amounts up to about 10%;preferably from about 2% to about 5% by weight of the pharmaceuticalcomposition.

Lubricating agents which may be used when desired in the instantcompositions include silicone oils or fluids such as substituted andunsubstituted polysiloxanes, e.g., dimethyl polysiloxane, also known asdimethicone. Other well known lubricating agents may be employed.

It is not expected that the inventive methods use compositions whichwill display significant adverse interactions with other synthetic ornaturally occurring substances. Thus, a compound of the presentinventive subject matter may be administered in combination with othercompounds and compositions useful for treating glioblastoma. Inparticular the inventive methods use compositions which may beadministered in combination with other inventive compositions, otherantineoplastic substances, and the like.

The optimal pharmaceutical formulations will be determined by oneskilled in the art depending upon considerations such as the route ofadministration and desired dosage. See, for example, “Remington'sPharmaceutical Sciences”, 18th ed. (1990, Mack Publishing Co., Easton,Pa. 18042), pp. 1435-1712, which is hereby incorporated by reference inits entirety. Such formulations may influence the physical state,stability, rate of in vivo release, and rate of in vivo clearance of thepresent therapeutic agents of the inventive subject matter.

Methods for Modulating Gene Expression

To identify genes that mediate pro-apoptotic response, global geneexpression changes were determined by oligonucleotide microarray(GeneChip® Human Genome U133A Affymetrix, Inc., containing 22,000 genes)in U87 cells. Data was collected at 30 minutes, 2 hours, and 24 hoursafter treatment with the inventive compositions in two independentexperiments. No consistent changes in gene expression were detectedafter 30 minutes of treatment with the inventive compositions. However,as shown in Table II, at 2 hours after treatment, Applicants identifiedseven genes that were up-regulated, five of them belonging to the heatshock protein family, and two that were down-regulated more thantwo-fold compared to DMSO-treated control cells. At 24 hours, theexpression of 84 genes was increased and of 65 genes was decreased. Theexpression of five genes influenced by treatment with the inventivecompositions has been confirmed by the real-time PCR analysis and isshown in Table III and IV.

Table II

At 2 hours after treatment with the inventive compositions, seven geneswere up-regulated and two were down-regulated more than 2-fold by thetreatment with the inventive compositions in two experiments.

Microarray Gene Gene Bank Fold change Gene name symbol number Exp1 Exp2heat shock 70 kDa protein 1A HSPA1A NM_005345 4.9 7.0 heat shock 70 kDaprotein 6 HSPA6 NM_002155 3.5 5.7 (HSP70B′) aldo-keto reductase family1, AKR1C2 U05598 4.9 5.3 member C2 (dihydrodiol dehydrogenase 2; bileacid binding protein; 3-alpha hydroxysteroid dehydrogenase, type III)heat shock 70 kDa protein 1B HSPA1B NM_005346 3.5 4.9 heme oxygenase(decycling) 1 HMOX1 NM_002133 4.3 4.6 solute carrier family 7, (cationicSLC7A11 AB040875 2.3 2.3 amino acid transporter, y+ system) member 11DnaJ (Hsp40) homolog, DNAJB1 NM_006145 2.0 2.1 subfamily B, member 1dual specificity phosphatase 6 DUSP6 BC003143 −2.5 −2.6 basichelix-loop-helix domain BHLHB2 NM_003670 −2.8 −3.7 containing, class B,2

Table III

At 24 hours after treatment with the inventive compositions, a total of149 differentially expressed genes with more than 2-fold change inexpression in both experiments were identified, the 84 which wereup-regulated are shown.

Microarray Fold Q-PCR Gene Gene Bank change Fold change Gene name symbolnumber Exp1 Exp2 Exp1 Exp2 Genes that increased in expression aftertreatment interleukin 1, beta IL1B NM_000576 7.7 27.9 N.D. 49.8 hemeoxygenase (decycling) 1 HMOX1 NM_002133 8.6 11.3 19.6 14.4 interleukin1, alpha IL1A M15329 4.3 8.0 15.6 11.2 colony stimulating factor 3 CSF3NM_000759 5.7 8.0 N.D. N.D. (granulocyte) leukemia inhibitory factor LIFNM_002309 7.0 4.0 N.D. N.D. (cholinergic differentiation factor) serine(or cysteine) proteinase SERPINB2 NM_002575 3.7 7.0 N.D. N.D. inhibitor,clade B (ovalbumin), member 2 hypothetical protein FLJ22761 FLJ22761NM_025130 3.5 5.7 N.D. N.D. neuregulin 1 NRG1 NM_013959 3.5 4.0 N.D.N.D. aldo-keto reductase family 1, AKR1C2 U05598 4.6 3.0 19.2 15.2member C2 (dihydrodiol dehydrogenase 2; bile acid binding protein;3-alpha hydroxysteroid dehydrogenase, type III) synovial sarcoma, Xbreakpoint 1 SSX1 NM_005635 2.0 11.3 N.D. N.D. interleukin 8 IL8AF043337 2.5 8.0 N.D. N.D. epiregulin EREG NM_001432 3.0 8.0 N.D. N.D.hepatitis delta antigen- DIPA BE966372 2.6 7.0 N.D. N.D. interactingprotein A dual specificity phosphatase 6 DUSP6 BC005047 2.6 6.5 N.D.N.D. kynureninase (L-kynurenine KYNU BC000879 2.3 4.6 N.D. N.D.hydrolase) dual specificity phosphatase 6 DUSP6 BC003143 3.0 4.0 N.D.N.D. interleukin 8 IL8 NM_000584 2.0 4.0 N.D. N.D. chemokine (C—X—Cmotif) CXCL3 NM_002090 2.8 3.7 N.D. N.D. ligand 3 dual specificityphosphatase 6 DUSP6 BC003143 2.6 3.5 N.D. N.D. kynureninase(L-kynurenine KYNU NM_003937 2.3 3.5 N.D. N.D. hydrolase) chromosome 8open reading C8orf4 NM_020130 2.3 3.2 N.D. N.D. frame 4 chemokine (C—X—Cmotif) CXCL2 M57731 2.3 3.2 N.D. N.D. ligand 2 hypothetical proteinFLJ13105 FLJ13105 NM_025001 2.6 3.2 N.D. N.D. sequestosome 1 SQSTM1N30649 2.8 3.0 N.D. N.D. phosphoprotein regulated by C8FW NM_025195 3.03.0 N.D. N.D. mitogenic pathways aldo-keto reductase family 1, AKR1C2BF508244 4.6 3.0 N.D. N.D. member C2 (dihydrodiol dehydrogenase 2; bileacid binding protein; 3-alpha hydroxysteroid dehydrogenase, type III)aldo-keto reductase family 1, AKR1C1 S68290 2.5 3.0 N.D. N.D. member C1(dihydrodiol dehydrogenase 1; 20-alpha (3- alpha)-hydroxysteroiddehydrogenase) spermidine/spermine N1- SAT M55580 2.3 3.0 N.D. N.D.acetyltransferase pregnancy-induced growth OKL38 NM_013370 3.7 3.0 N.D.N.D. inhibitor diphtheria toxin receptor DTR NM_001945 2.0 2.8 N.D. N.D.(heparin-binding epidermal growth factor-like growth factor) immediateearly response 3 IER3 NM_003897 2.1 2.8 N.D. N.D. DEAD (Asp-Glu-Ala-Asp)box DDX10 NM_004398 3.2 2.6 N.D. N.D. polypeptide 10 solute carrierfamily 3 SLC3A2 NM_002394 2.0 2.6 N.D. N.D. (activators of dibasic andneutral amino acid transport), member 2 NA NA AA488687 2.5 2.6 N.D. N.D.NA NA M27830 24.3 2.5 N.D. N.D. spermidine/spermine N1- SAT NM_0029702.1 2.5 N.D. N.D. acetyltransferase BCL2-associated athanogene BAG1AF116273 2.3 2.5 N.D. N.D. kynureninase (L-kynurenine KYNU BC000879 4.32.5 N.D. N.D. hydrolase) cAMP responsive element CREM D14826 2.1 2.5N.D. N.D. modulator aldo-keto reductase family 1, AKR1C2 M33376 2.0 2.5N.D. N.D. member C2 (dihydrodiol dehydrogenase 2; bile acid bindingprotein; 3-alpha hydroxysteroid dehydrogenase, type III) v-mafmusculoaponeurotic MAFF AL021977 2.0 2.5 N.D. N.D. fibrosarcoma oncogeneuroporphyrinogen UROD AW614435 2.6 2.5 N.D. N.D. decarboxylase solutecarrier family 25 SLC25A6 AI961224 2.1 2.5 N.D. N.D. (mitochondrialcarrier; adenine nucleotide translocator), member 6 cathepsin L CTSLNM_001912 2.3 2.5 N.D. N.D. ornithine decarboxylase 1 ODC1 NM_002539 2.12.5 N.D. N.D. dual specificity phosphatase 1 DUSP1 NM_004417 2.5 2.5N.D. N.D. glucosaminyl (N-acetyl) GCNT3 NM_004751 5.3 2.5 N.D. N.D.transferase 3, mucin type galactosidase, alpha GLA NM_000169 2.3 2.3N.D. N.D. hypothetical protein FLJ20489 FLJ20489 H14241 2.6 2.3 N.D.N.D. deleted in liver cancer 1 DLC1 AF026219 2.5 2.3 N.D. N.D.thioredoxin reductase 1 TXNRD1 NM_003330 2.3 2.3 N.D. N.D.glutamate-cysteine ligase, GCLM NM_002061 2.1 2.3 N.D. N.D. modifiersubunit NAD(P)H dehydrogenase, NQO1 AI039874 2.3 2.3 N.D. N.D. quinone 1GABA(A) receptors associated GABARAPL3 AF180519 3.2 2.3 N.D. N.D.protein like 3 decay accelerating factor for DAF NM_000574 3.0 2.3 N.D.N.D. complement (CD55, Cromer blood group system) aminolevulinate,delta-, ALAS1 NM_000688 2.6 2.3 N.D. N.D. synthase 1prostaglandin-endoperoxide PTGS2 NM_000963 2.1 2.3 N.D. N.D. synthase 2(prostaglandin G/H synthase and cyclooxygenase) nuclear receptorinteracting NRIP1 AI824012 2.0 2.3 N.D. N.D. protein 1 retinoic acidinduced 3 RAI3 NM_003979 2.0 2.3 N.D. N.D. synovial sarcoma, Xbreakpoint 1 SSX1 BC001003 2.1 2.1 N.D. N.D. hypothetical proteinLOC54103 LOC54103 AK026747 2.1 2.1 N.D. N.D. NA NA M11167 4.0 2.1 N.D.N.D. C2f protein C2F U72514 2.3 2.1 N.D. N.D. prostaglandin E synthasePTGES AF010316 2.6 2.1 N.D. N.D. synovial sarcoma, X breakpoint 3 SSX3X79200 2.3 2.1 N.D. N.D. tetratricopeptide repeat domain 4 TTC4NM_004623 2.3 2.1 N.D. N.D. annexin A10 ANXA10 AF196478 2.3 2.1 N.D.N.D. synovial sarcoma, X breakpoint 2 SSX2 AF257500 2.1 2.0 N.D. N.D.TRAF and TNF receptor TTRAP NM_016614 2.5 2.0 N.D. N.D. associatedprotein tripartite motif-containing 16 TRIM16 NM_006470 2.3 2.0 N.D.N.D. a disintegrin-like and ADAMTS5 NM_007038 2.5 2.0 N.D. N.D.metalloprotease (reprolysin type) with thrombospondin type 1 motif, 5(aggrecanase-2) NA NA NM_014903 2.0 2.0 N.D. N.D. jumonji homolog(mouse) JMJ NM_004973 2.5 2.0 N.D. N.D. peptidylprolyl isomerase F PPIFBC005020 2.1 2.0 N.D. N.D. (cyclophilin F) piccolo (presynapticcytomatrix PCLO AB011131 2.0 2.0 N.D. N.D. protein) sphingosine kinase 1SPHK1 NM_021972 2.6 2.0 N.D. N.D. solute carrier family 7, SLC7A11AB040875 2.3 2.0 N.D. N.D. (cationic amino acid transporter, y+ system)member 11 NAD(P)H dehydrogenase, NQO1 BC000906 2.0 2.0 N.D. N.D. quinone1 tetratricopeptide repeat domain 4 TTC4 W22690 2.3 2.0 N.D. N.D.nuclear receptor interacting NRIP1 NM_003489 2.1 2.0 N.D. N.D. protein 1glutamate-cysteine ligase, GCLC NM_001498 2.1 2.0 N.D. N.D. catalyticsubunit

Table IV

At 24 hours after treatment with the inventive compositions, a total of149 differentially expressed genes with more than 2-fold change inexpression in both experiments were identified, the 65 which weredown-regulated are shown.

Microarray Q-PCR Fold Fold change change Gene name Gene symbol Gene Banknumber Exp1 Exp2 Exp1 Exp2 Genes that decreased in expression aftertreatment fatty acid binding FABP3 AI041520 −3.7 17.1 N.D. N.D. protein3, muscle and heart (mammary- derived growth inhibitor) collagen, typeI, COL1A2 NM_000089 −3.5 −4.9 N.D. N.D. alpha 2 single-stranded DNASSBP2 NM_012446 −3.5 −4.6 −10.6 −3.2 binding protein 2 lysyloxidase-like 1 LOXL1 NM_005576 −4.3 −4.3 N.D. N.D. leucine rich repeatLRRC17 NM_005824 −3.5 −4.0 N.D. N.D. containing 17 thioredoxin TXNIPAI439556 −4.3 −3.5 N.D. N.D. interacting protein chromosome 5 openC5orf13 NM_004772 −3.7 −3.5 N.D. N.D. reading frame 13 carbonicanhydrase CA12 BC000278 −3.2 −2.0 N.D. N.D. XII Tara-like proteinHRIHFB2122 AF281030 −2.6 −2.0 N.D. N.D. N-myc downstream NDRG1 NM_006096−2.3 −2.0 N.D. N.D. regulated gene 1 hypothetical protein FLJ20151BC001012 −2.1 −2.0 N.D. N.D. FLJ20151 collagen, type V, COL5A1 AI130969−2.6 −2.0 N.D. N.D. alpha 1 tumor necrosis factor TNFRSF9 BC006196 −2.3−2.0 N.D. N.D. receptor superfamily, member 9 NA NA NM_017635 −2.1 −2.0N.D. N.D. stearoyl-CoA SCD AB032261 −2.0 −2.0 N.D. N.D. desaturase(delta-9- desaturase) collagen, type V, COL5A1 AI983428 −2.6 −2.0 N.D.N.D. alpha 1 nuclear receptor NR2F2 AL554245 −2.6 −2.0 N.D. N.D.subfamily 2, group F, member 2 old astrocyte OASIS BG328407 −6.1 −2.1N.D. N.D. specifically induced substance inhibitor of DNA ID2 NM_002166−3.2 −2.1 N.D. N.D. binding 2, dominant negative helix-loop- helixprotein artemin ARTN NM_003976 −2.8 −2.1 N.D. N.D. chondroitin sulfateCSPG2 D32039 −2.8 −2.1 N.D. N.D. proteoglycan 2 (versican) S100 calciumbinding S100A3 NM_002960 −2.1 −2.1 N.D. N.D. protein A3 stearoyl-CoA SCDAF116616 −14.9 −2.1 N.D. N.D. desaturase (delta-9- desaturase) WNT1inducible WISP1 NM_003882 −2.1 −2.1 N.D. N.D. signaling pathway protein1 solute carrier family SLC2A1 NM_006516 −3.2 −2.1 N.D. N.D. 2(facilitated glucose transporter), member 1 KIAA0876 protein KIAA0876AW237172 −2.5 −2.1 N.D. N.D. protocadherin gamma PCDHGA3 AF152509 −21.1−2.3 N.D. N.D. subfamily A, 3 matrilin 2 MATN2 NM_002380 −2.1 −2.3 N.D.N.D. artemin ARTN AF115765 −3.0 −2.3 N.D. N.D. intermediate filamentSYNCOILIN NM_030786 −2.3 −2.3 N.D. N.D. protein syncoilin paternallyexpressed PEG10 BE858180 −6.1 −2.3 N.D. N.D. 10 hypothetical proteinFLJ20716 NM_017938 −2.3 −2.3 N.D. N.D. FLJ20716 insulin-like growthIGFBP3 BF340228 −5.3 −2.3 N.D. N.D. factor binding protein 3hypothetical protein FLJ10970 NM_018286 −2.3 −2.3 N.D. N.D. FLJ10970chromosome 14 open C14orf132 NM_020215 −2.0 −2.3 N.D. N.D. reading frame132 lanosterol synthase LSS AW084510 −2.3 −2.5 N.D. N.D.(2,3-oxidosqualene- lanosterol cyclase) KIAA0367 protein KIAA0367AB002365 −2.3 −2.5 N.D. N.D. hypothetical protein LOC284266 AK025833−3.0 −2.5 N.D. N.D. LOC284266 calcium/calmodulin- CaMKIINalpha NM_018584−2.1 −2.5 N.D. N.D. dependent protein kinase II dudulin2 TSAP6 NM_018234−2.3 −2.5 N.D. N.D. 3-hydroxy-3- HMGCS1 BG035985 −2.1 −2.5 N.D. N.D.methylglutaryl- Coenzyme A synthase 1 (soluble) collagen, type V, COL5A1N30339 −2.0 −2.5 N.D. N.D. alpha 1 nuclear receptor NR2F2 AV703465 −2.1−2.5 N.D. N.D. subfamily 2, group F, member 2 kinesin family KIF20ANM_005733 −2.5 −2.5 N.D. N.D. member 20A paternally expressed PEG10AL582836 −2.5 −2.5 N.D. N.D. 10 growth associated GAP43 NM_002045 −2.5−2.5 N.D. N.D. protein 43 lanosterol synthase LSS D63807 −6.5 −2.5 N.D.N.D. (2,3-oxidosqualene- lanosterol cyclase) papillomavirus PBFNM_018660 −3.0 −2.6 N.D. N.D. regulatory factor PRF-1 hypotheticalprotein DKFZp434K NM_017606 −2.6 −2.8 N.D. N.D. DKFZp434K1210 1210thioredoxin TXNIP NM_006472 −5.3 −2.8 N.D. N.D. interacting proteinchromosome 14 open C14orf139 NM_024633 −2.3 −2.8 N.D. N.D. reading frame139 hypothetical protein LOC283824 AI693140 −2.0 −2.8 N.D. N.D.LOC283824 fatty acid binding FABP3 NM_004102 −3.5 −3.0 N.D. N.D. protein3, muscle and heart (mammary- derived growth inhibitor) collagen, typeI, COL1A1 AI743621 −2.8 −2.8 N.D. N.D. alpha 1 collagen, type I, COL1A1K01228 −3.7 −3.0 N.D. N.D. alpha 1 protease, serine, 23 SPUVE NM_007173−2.3 −3.0 N.D. N.D. thioredoxin TXNIP AA812232 −9.8 −3.2 N.D. N.D.interacting protein guanylate binding GBP1 NM_002053 −2.5 −3.2 N.D. N.D.protein 1, interferon- inducible, 67 kDa inhibitor of DNA ID3 NM_002167−2.0 −3.2 N.D. N.D. binding 3, dominant negative helix-loop- helixprotein old astrocyte OASIS AF055009 −2.5 −3.5 N.D. N.D. specificallyinduced substance synaptopodin SYNPO NM_007286 −3.0 −3.7 N.D. N.D.DKFZP586A0522 DKFZP586A NM_014033 −3.0 −3.7 N.D. N.D. protein 0522single-stranded DNA SSBP2 AF077048 −2.6 −3.7 N.D. N.D. binding protein 2leucine rich repeat LRRN3 AI221950 −2.6 −4.6 N.D. N.D. neuronal 3hepatitis delta DIPA NM_006848 −2.3 −12.1 N.D. N.D. antigen-interactingprotein A

Surprisingly, two known pro-inflammatory genes, IL-1α and IL-1β, weredramatically up-regulated in expression upon treatment with theinventive compositions. Upon up-regulation of heat shock protein familygenes, the inventive compositions would be expected to produce a strongprotective response in treated glioblastoma cells. But clearly any suchresponse produced is insufficient to protect those cells from apoptosis.Thus, it is not clear whether induction of the protective heat shockcellular mechanism is necessary for the efficient induction of anapoptotic response, or whether the GBM cell killing would be enhanced ifthis response is prevented or diminished. In any event, Applicantsexpect that some or all of the genes up-regulated in response totreatment with the inventive compositions, individually or collectively,induce apoptosis.

Without being bound to a particular mechanism of action, the obtainedresults suggest that phytonutrients, which are present in the herbalpreparation of the inventive compositions, modulate the expression ofgenes involved in regulation of apoptosis and inflammation, andindividually or collectively induce apoptosis in glioblastoma cells.

Thus, the inventive subject matter further relates to methods formodulating gene expression in a target cell of one or more genesselected from the group consisting of: interleukin-1α, interleukin-1β,heme oxygenase 1, aldo-keto reductase family 1, member C2, colonystimulating factor 3, leukemia inhibitory factor, heat shock 70 kDaprotein, and combinations thereof, comprising administration of aneffective amount of a composition comprising therapeutically effectiveamounts of supercritical extracts of rosemary, turmeric, oregano andginger; and therapeutically effective amounts of hydroalcoholic extractsof holy basil, ginger, turmeric, Scutellaria baicalensis, rosemary,green tea, huzhang, Chinese goldthread, and barberry.

In one aspect of the inventive subject matter, said composition isadministered orally.

In another aspect of the inventive subject matter, said orallyadministered composition is in the form of one or more capsules, one ormore tablets, or one or more pills.

In a preferred embodiment, said composition comprises:

-   -   (A) from about 4.5% to about 7.5% by weight of the        hydroalcoholic extract of ginger;    -   (B) from about 5.5% to about 8.5% by weight of the supercritical        extract of ginger;    -   (C) from about 1.0% to about 1.5% by weight of the supercritical        extract of turmeric;    -   (D) from about 10.0% to about 16.0% by weight of the        supercritical extract of rosemary;    -   (E) from about 4.0% to about 6.0% by weight of the supercritical        extract of oregano;    -   (F) from about 10.0% to about 16.0% by weight of the        hydroalcoholic extract of turmeric;    -   (G) from about 5.5% to about 8.0% by weight of the        hydroalcoholic extract of rosemary;    -   (H) from about 10.0% to about 16.0% by weight of the        hydroalcoholic extract of holy basil;    -   (I) from about 10.0% to about 16.0% by weight of the        hydroalcoholic extract of green tea;    -   (J) from about 8.0% to about 12.0% by weight of the        hydroalcoholic extract of huzhang;    -   (K) from about 4.0% to about 6.0% by weight of the        hydroalcoholic extract of Chinese goldthread;    -   (L) from about 4.0% to about 6.0% by weight of the        hydroalcoholic extract of barberry; and    -   (M) from about 2.0% to about 3.0% by weight of the        hydroalcoholic extract of Scutellaria baicalensis.

It is expected that one or more of the proteins described hereincontrols, or contributes to controlling, cellular apoptosis. Withoutbeing bound to a particular mechanism of action, it is further expectedthat by administering an inventive composition, alone or in combinationwith other therapeutic compounds and compositions, the apoptotic processis up-regulated, resulting in cell death of target cell line(s), such ascancer cells, and in particular GBM cells.

Heme oxygenase (decycling) 1 (hereinafter “HMOX1”) is an inducibleenzyme of the heat shock protein family that is essential in hemecatabolism, where it cleaves heme to form biliverdin. It is one of thekey cellular defensive enzymes, is involved in regulation of theI-kappaB kinase/NF-kappaB cascade, has signal transducer activity andoxidoreductase activity, and plays a protective role as an antioxidantin the lung and antagonizes inflammation in an animal skin wound model(see Wagener F A, van Beurden H E, von den Hoff J W et al: The heme-hemeoxygenase system: a molecular switch in wound healing. Blood102:521-528, 2003). Its role in GBM is presently unknown, but Applicantsexpect it to be important because it appears to plays a role inregulation of apoptosis. As shown in FIG. 2, administration of theinventive compositions up-regulates expression of HMOX1 protein in U87GBM cell line after 24 hours of stimulation.

Aldo-keto reductase family 1, member C2 (hereinafter “AKR1C2”) is amember of the aldo-oxo reductase superfamily that has been implicated inthe metabolism of glucocorticoids, progestins, bile acid precursors,COX-2-derived prostaglandins and xenobiotic carbonyl compounds and thusApplicants expect it to be important in GBM tumorigenesis. Incomparison, in prostate cells it prevents activation of androgenreceptor (see Rizner T L, Lin H K, Peehl D M et al: Human type 33-alpha-hydroxysteroid dehdydrogenase (aldo-keto reductase 1 C2) andandrogen metabolism in prostate cells. Endocrinol, 144:2922-2932, 2003).

Heat shock 70 kDa protein 1A (hereinafter “HSPA1A”), heat shock 70 kDaprotein 1B (hereinafter “HSPA1B”), and heat shock 70 kDa protein 6(hereinafter “HSPA6”) are members of the heat shock family of induciblegenes, and which appear to be involved in response to heat shock andother physiological challenges, protein folding and molecularchaperoning, mRNA catabolism, and ATP binding.

Interleukin-1A (hereinafter “IL1A”) gene encodes IL-1α protein, which isa member of the interleukin 1 cytokine family. This cytokine is apleiotropic cytokine involved in various immune responses, inflammatoryprocesses, and hematopoiesis. This cytokine is produced by monocytes andmacrophages as a proprotein, which is proteolytically processed andreleased in response to cell injury, and thus induces apoptosis.

Interleukin-1B (hereinafter “IL1B”) gene encodes IL-1β protein, which isa member of the interleukin 1 cytokine family. This cytokine is producedby activated macrophages as a proprotein, which is proteolyticallyprocessed to its active form by caspase 1 (CASP1/ICE). This cytokine isan important mediator of the inflammatory response, and is involved in avariety of cellular activities, including cell proliferation,differentiation, and apoptosis. The induction of cyclooxygenase-2(PTGS2/COX2) by this cytokine in the central nervous system (CNS) isfound to contribute to inflammatory pain hypersensitivity.

Colony stimulating factor 3 (hereinafter “CSF3”) gene encodes colonystimulating factor 3 protein, which is a cytokine that controls theproduction, differentiation, and function of granulocytes. The activeprotein is found extracellularly.

Leukemia inhibitory factor (hereinafter “LIF”) gene encodes leukemiainhibitory factor protein, which is a cytokine that induces macrophagedifferentiation. Neurotransmitters and neuropeptides, well known fortheir role in the communication between neurons, are also capable ofactivating monocytes and macrophages and inducing chemotaxis in immunecells.

Solute carrier family 7, (cationic amino acid transporter, y+ system)member 11 (hereinafter “SLC7A11”) is a member of a heteromericNa(+)-independent anionic amino acid transport system highly specificfor cystine and glutamate. In addition to cystine:glutamate antiporteractivity, SLC7A11 is involved in protein complex assembly, amino acidtransport, and has amino acid permease activity.

DnaJ homolog, subfamily B, member 1 (hereinafter “DNAJB1”), also knownas Hsp40, is a member of the heat shock family of inducible genes, andwhich appears to be involved in response to heat shock and otherphysiological challenges, protein folding, and molecular chaperoning.

Dual specificity phosphatase 6 (hereinafter “DUSP6”) is a proteintyrosine phosphatase which appears to be involved in cell cycleregulation, particularly regulation of mitogen-activated protein kinase,protein amino acid dephosphorylation, and apoptosis. This phosphataseinactivates its target kinase(s) by dephosphorylating both thephosphoserine/threonine and phosphotyrosine residues. DUSP6 has proteinserine/threonine phosphatase activity, protein tyrosine phosphataseactivity, hydrolase activity, and MAP kinase phosphatase activity.

Basic helix-loop-helix domain containing, class B, 2 (hereinafter“BHLHB2”) encodes a basic helix-loop-helix protein expressed in varioustissues. Expression in the chondrocytes is responsive to the addition ofBt2cAMP and is believed to be involved in the control of celldifferentiation. BHLHB2 is involved in DNA-dependent regulation oftranscription and has transcription factor activity.

Route(s) of Administration

The compounds and compositions are preferably administered orally in theform of capsules, tablets, aqueous suspensions, or solutions. Tabletsmay contain carriers such as lactose and corn starch, and/or lubricatingagents such as magnesium stearate. Capsules may contain diluentsincluding lactose and dried corn starch. Aqueous suspensions may containemulsifying and suspending agents combined with the active ingredient.The oral dosage forms may further contain sweetening, flavoring,coloring agents, or combinations thereof. Delivery in an entericallycoated tablet, caplet, or capsule, to further enhance stability andprovide release in the intestinal tract to improve absorption, is thebest mode of administration currently contemplated.

Dosage

Dosage levels on the order of about 0.001 mg to about 100 mg perkilogram body weight of the active ingredient compounds or compositionsare useful in the treatment of the above conditions, with preferredlevels ranging from 200 mg per day to 1600 mg per day. The compounds andcompositions of the present inventive subject matter may usually begiven in two or three doses daily. Starting with a low dose (200-300 mg)twice daily and slowly working up to higher doses if needed is apreferred strategy. The amount of active ingredient that may be combinedwith the carrier materials to produce a single dosage form will varydepending upon the host treated and the particular mode ofadministration.

It is understood, however, that a specific dose level for any particularpatient will depend upon a variety of factors, including the activity ofthe specific compound employed; the age, body weight, general health,sex and diet of the patient; the time of administration; the rate ofexcretion; drug combination; the severity of the particular disorderbeing treated; and the form of administration. One of ordinary skill inthe art would appreciate the variability of such factors and would beable to establish specific dose levels using no more than routineexperimentation.

EXAMPLES

The following examples are illustrative of the present inventive subjectmatter and are not intended to be limitations thereon. Unless otherwiseindicated, all percentages are based upon 100% by weight of the finalcomposition.

Example 1 Preparation of the Inventive Compositions

The inventive compositions are prepared by methods known in the art, anddisclosed in Applicants' U.S. Pat. No. 6,387,416. The preparation of thecomponent elements of the inventive compositions is summarized asfollows:

The hydroalcoholic extract of ginger used in the inventive compositionsis preferably prepared as follows. The ginger rhizome, which ispreferably cryogenically ground to preserve heat sensitive components,is subjected to supercritical extraction to obtain: (i) an oil extract,referred to herein as “the supercritical extract” of ginger, containingdelicate lipophilic components, and (ii) an oil-free residue. Theoil-free residue is then extracted in a water/alcohol, preferablywater/ethanol, mixture composed of 60-80 parts alcohol and 40-20 partswater. The alcohol/water liquid is then evaporated off, leaving apowdered extract residue, referred to herein as “the hydroalcoholicextract” of ginger.

The composition of this invention will preferably contain thesupercritical extract and the hydroalcoholic extract of ginger at aweight ratio of preferably from about 0.9 to about 1.4 parts, morepreferably from about 1.1 to about 1.3 parts, most preferably about 1.17parts, of supercritical extract per 1 part post-supercriticalhydroalcoholic extract.

The supercritical extracts of ginger, rosemary, turmeric and oreganoused in the inventive compositions can be prepared according to knownsupercritical extraction methods, such as disclosed, e.g., in E. Stahl,K. W. Quirin, D. Gerard, Dense Gases for Extraction and Refining,Springer Verlag 4 1988, which is hereby incorporated by referenceherein.

The hydroalcoholic extracts of rosemary, turmeric, holy basil, greentea, huzhang, Chinese goldthread, barberry and Scutellaria baicalensisused in the inventive compositions can be prepared according toconventional hydroalcoholic extraction techniques. For example, thehydroalcoholic extracts can be prepared by extracting the plant portionin a water/alcohol (preferably water/ethanol) mixture (preferablycomposed of 60-80 parts alcohol and 40-20 parts water), and thenevaporating off the water/alcohol liquid, leaving a powdered extractresidue (referred to herein as “the hydroalcoholic extract”).

In the composition of this invention, the hydroalcoholic extract ofturmeric and the supercritical extract of turmeric will preferably bepresent at a weight ratio of preferably from about 8 to about 12 parts,more preferably from about 9 parts to about 11 parts, most preferablyabout 10 parts, of hydroalcoholic extract per 1 part of supercriticalextract.

The composition of this invention will preferably contain thesupercritical extract of rosemary and the hydroalcoholic extract ofrosemary at a weight ratio of preferably from about 1.6 to about 2.4parts, more preferably from about 1.8 to about 2.2 parts, mostpreferably about 2.0 parts, of supercritical extract per 1 part ofhydroalcoholic extract.

The hydroalcoholic extract of ginger used in the inventive compositionswill preferably contain from about 2.4% to about 3.6%, more preferablyfrom about 2.7% to about 3.3%, most preferably about 3.0%, by weight ofpungent compounds (e.g., shogaol).

The supercritical extract of ginger used in the inventive compositionswill contain preferably from about 24% to about 36%, more preferablyfrom about 27% to about 33%, most preferably about 30%, by weight ofpungent compounds (e.g., shogaol) and preferably from about 6.4% toabout 9.6%, more preferably from about 7.2% to about 8.8%, mostpreferably about 8%, by weight of zingiberene.

The supercritical extract of turmeric used in the inventive compositionswill contain preferably from about 36% to about 54%, more preferablyfrom about 40.5% to about 49.5%, most preferably about 45%, by weight ofturmerones.

The supercritical extract of rosemary used in the inventive compositionswill contain preferably from about 18.4% to about 27.6%, more preferablyfrom about 20.7% to about 25.3%, most preferably about 23%, by weight oftotal phenolic antioxidants (“TPA”).

The supercritical extract of oregano used in the inventive compositionswill contain preferably from about 0.64% to about 0.96%, more preferablyfrom about 0.72% to about 0.88%, most preferably about 0.8%, by weightof TPA.

The hydroalcoholic extract of turmeric used in the inventivecompositions will contain preferably from about 5.6% to about 8.4%, morepreferably from about 6.3% to about 7.7%, most preferably about 7%, byweight of curcumin.

The hydroalcoholic extract of rosemary used in the inventivecompositions will contain preferably from about 18.4% to about 27.6%,more preferably from about 20.7% to about 25.3%, most preferably about23%, by weight of TPA.

The hydroalcoholic extract of holy basil used in the inventivecompositions will contain preferably from about 1.6% to about 2.4%, morepreferably from about 1.8% to about 2.2%, most preferably about 2%, byweight of ursolic acid.

The hydroalcoholic extract of green tea used in the inventivecompositions will contain preferably from about 36% to about 54%, morepreferably from about 40.5% to about 49.5%, most preferably about 45%,by weight of polyphonies.

The hydroalcoholic extract of huzhang used in the inventive compositionswill contain preferably from about 6.4% to about 9.6%, more preferablyfrom about 7.2% to about 8.8%, most preferably about 8%, by weight ofresveratrol.

The hydroalcoholic extract of Chinese goldthread used in the inventivecompositions will contain preferably from about 4.8% to about 7.2%, morepreferably from about 5.4% to about 6.6%, most preferably about 6%, byweight of berberine.

The hydroalcoholic extract of barberry used in the inventivecompositions will contain preferably from about 4.8% to about 7.2%, morepreferably from about 5.4% to about 6.6%, most preferably about 6%, byweight of berberine.

Example 2 Effect of the Inventive Compositions on Glioblastoma Cells

The inventive compositions, diluted in DMSO at 0.1 μl/ml, inducedapoptosis of U87 cells compared to olive oil/DMSO control at 48 hours(55.3% vs. 1.7%) as determined by TUNEL analysis. These results, asshown in FIG. 2, demonstrate that the inventive compositions stronglysuppress U87 cells and induce apoptosis. Exponentially grown U87 cellsin DMEM with 1% FBS were treated with Zyflamend or olive oil at finalconcentration of 0.1 microl/ml, or left untreated. After 24 hours,floating and attached cells were collected, fixed in 1% paraformaldehydein phosphate buffered saline (PBS), washed in PBS, resuspended in 70%ethanol and analyzed by flow cytometry. The APO-BRDU kit, a two colorstaining method for labeling DNA breaks and total cellular DNA was usedto detect apoptotic cells. Percentage of apoptotic cells is indicated inthe upper right corner of each panel of FIG. 2.

Example 3 Effect of the Inventive Compositions on Gene Expression

In two independent experiments, total RNA was isolated from U87 cells at2 hours and 24 hours after treatment with the inventive compositions andin vitro transcripts were hybridized to GeneChip® Human Genome U133AAffymetrix oligonucleotide microarray. In total, at 2 hours aftertreatment with the inventive compositions, Applicants identified 7 genesthat were up-regulated, five of which belong to the heat shock proteinfamily, and two genes that were down-regulated more than 2-fold in bothexperiments compared to the DMSO-treated control cells. At 24 hours, theexpression of 84 genes was increased and of 65 genes was decreased. Theexpression of eight genes influenced by treatment with the inventivecompositions has been confirmed by the real-time PCR analysis andimmunoblotting evaluation is in progress.

Unexpectedly, the expression of interleukin-1

and interleukin-1

genes, potent mediators of inflammation and immunity, was significantlyinduced, up to 30-fold, at 24 hours after treatment with the inventivecompositions of U87 cells. The IL-1 cytokine is known to be produced inresponse to cell injury, but it remains to be demonstrated that it playsa role in induction of apoptosis by the inventive compositions in U87glioblastoma cells. Regardless of the mechanism of action, the obtainedresults suggest that naturally occurring COX-2 and 5-LO inhibitors fromthe inventive herbal preparation modulate the expression of functionallydiverse groups of genes, including those involved in regulation ofapoptosis and inflammation, that collectively induce apoptosis inglioblastoma cell line U87.

Example 4 Effect of the Inventive Compositions on Pre-MalignantGlioblastoma

A patient presents for treatment of a low-grade or high-grade neoplasmof the central nervous system. An inventive composition is administeredto said patient over a course of treatment lasting for several weeks,resulting in no significant side effects. The patient experiences areversal in the growth of neoplastic cells and death of existingneoplastic cells, resulting in the neoplasia becoming undetectable.

Example 5 Effect of the Inventive Compositions on Glioblastoma

A patient presents for treatment of a malignant grade IV glioblastoma ofthe central nervous system, confirmed by manual examination and biopsyof the tumor. An inventive composition is administered to said patientover a course of treatment lasting for several months, resulting in nosignificant side effects. The patient experiences a reversal in thegrowth rate of tumor cells, death of existing tumor cells and reductionin tumor size, and no metastasis of the tumor. With continuingtreatment, the patient continues to exhibit no secondary symptoms ofglioblastoma, no long term side effects of the treatment, and nometastasis of the tumor.

Example 6 Up-Regulation of HMOX-1

Protein extracts were prepared from human U87 GBM cells treated with theinventive compositions (“ZYF”), DMSO control (“DMSO”), or non-treated(“NT”) cells at 2 and 24 hours after stimulation. Immunoblotting withanti-Heme Oxygenase-1 (HMOX-1) monoclonal antibodies obtained fromStressgen Biotechnology, Canada was done according to recommendedprotocol. As shown in FIG. 2, administration of the inventivecompositions up-regulates expression of HMOX-1 protein in U87 GBM cellline after 24 hours of stimulation.

The inventive subject matter being thus described, it will be obviousthat the same may be modified or varied in many ways. Such modificationsand variations are not to be regarded as a departure from the spirit andscope of the inventive subject matter and all such modifications andvariations are intended to be included within the scope of the followingclaims.

1. A method for increasing gene expression, to induce cellular apoptosisin a target glioblastoma cell of a subject in need thereof, of one ormore genes selected from the group consisting of: interleukin-1α,interleukin-1β, heme oxygenase 1, aldo-keto reductase family 1, memberC2, colony stimulating factor 3, leukemia inhibitory factor, heat shock70 kDa protein 1A, heat shock 70 kDa protein 1B, heat shock 70 kDaprotein 6, and combinations thereof, comprising administering aneffective amount of a composition comprising therapeutically effectiveamounts of supercritical extracts of rosemary, turmeric, oregano andginger; and therapeutically effective amounts of hydroalcoholic extractsof holy basil, ginger, turmeric, Scutellaria baicalensis, rosemary,green tea, huzhang, Chinese goldthread, and barberry.
 2. The method ofclaim 1, wherein said composition is administered orally.
 3. The methodof claim 2, wherein the orally administered composition is in the formof one or more capsules, one or more tablets, or one or more pills. 4.The method of claim 1, wherein the composition comprises: a. from about4.5% to about 7.5% by weight of the hydroalcoholic extract of ginger; b.from about 5.5% to about 8.5% by weight of the supercritical extract ofginger; c. from about 1.0% to about 1.5% by weight of the supercriticalextract of turmeric; d. from about 10.0% to about 16.0% by weight of thesupercritical extract of rosemary; e. from about 4.0% to about 6.0% byweight of the supercritical extract of oregano; f. from about 10.0% toabout 16.0% by weight of the hydroalcoholic extract of turmeric; g. fromabout 5.5% to about 8.0% by weight of the hydroalcoholic extract ofrosemary; h. from about 10.0% to about 16.0% by weight of thehydroalcoholic extract of holy basil; i. from about 10.0% to about 16.0%by weight of the hydroalcoholic extract of green tea; j. from about 8.0%to about 12.0% by weight of the hydroalcoholic extract of huzhang; k.from about 4.0% to about 6.0% by weight of the hydroalcoholic extract ofChinese goldthread; l. from about 4.0% to about 6.0% by weight of thehydroalcoholic extract of barberry; and m. from about 2.0% to about 3.0%by weight of the hydroalcoholic extract of Scutellaria baicalensis. 5.The method of claim 1, wherein said subject is suffering from acancerous tumor of the central nervous system, and wherein the canceroustumor is glioblastoma.
 6. The method of claim 5, wherein the amount ofthe composition administered to said subject is effective to treatglioblastoma.
 7. The method of claim 1, wherein the weight ratio of thesupercritical extract of ginger to the hydroalcoholic extract of gingeris from about 0.9:1 to about 1.4:1.
 8. The method of claim 1, whereinthe weight ratio of the hydroalcoholic extract of turmeric to thesupercritical extract of turmeric is from about 8:1 to about 12:1. 9.The method of claim 1, wherein the weight ratio of the supercriticalextract of rosemary to the hydroalcoholic extract of rosemary is fromabout 1.6:1 to about 2.4:1.
 10. The method of claim 1, wherein thehydroalcoholic extract of ginger comprises from about 2.4% to about 3.6%by weight of pungent compounds.
 11. The method of claim 1, wherein thesupercritical extract of ginger comprises from about 24% to about 36% byweight of pungent compounds and from about 6.4% to about 9.6% by weightof zingiberene.
 12. The method of claim 1, wherein the supercriticalextract of turmeric comprises from about 36% to about 54% by weight ofturmerones.
 13. The method of claim 1, wherein the supercritical extractof rosemary comprises from about 18.4% to about 27.6% by weight of totalphenolic antioxidants.
 14. The method of claim 1, wherein thesupercritical extract of oregano comprises from about 0.64% to about0.96% by weight of total phenolic antioxidants.
 15. The method of claim1, wherein the hydroalcoholic extract of turmeric comprises from about5.6% to about 8.4% by weight of curcumin.
 16. The method of claim 1,wherein the hydroalcoholic extract of rosemary comprises from about18.4% to about 27.6% by weight of total phenolic antioxidants.
 17. Themethod of claim 1, wherein the hydroalcoholic extract of holy basilcomprises from about 1.6% to about 2.4% by weight of ursolic acid. 18.The method of claim 1, wherein the hydroalcoholic extract of green teacomprises from about 36% to about 54% by weight of polyphenols.
 19. Themethod of claim 1, wherein the hydroalcoholic extract of huzhangcomprises from about 6.4% to about 9.6% by weight of resveratrol. 20.The method of claim 1, wherein the hydroalcoholic extract of Chinesegoldthread comprises from about 4.8% to about 7.2% by weight ofberberine.
 21. The method of claim 1, wherein the hydroalcoholic extractof barberry comprises from about 4.8% to about 7.2% by weight ofberberine.
 22. The method of claim 1, wherein said composition providedin step (a) comprises: a. from about 4.5% to about 7.5% by weight of thehydroalcoholic extract of ginger, wherein the extract comprises fromabout 2.4% to about 3.6% by weight of pungent compounds; b. from about5.5% to about 8.5% by weight of the supercritical extract of ginger,wherein the extract comprises from about 24% to about 36% by weight ofpungent compounds and from about 6.4% to about 9.6% by weight ofzingiberene; c. from about 1.0% to about 1.5% by weight of thesupercritical extract of turmeric, wherein the extract comprises fromabout 36% to about 54% by weight of turmerones; d. from about 10.0% toabout 16.0% by weight of the supercritical extract of rosemary, whereinthe extract comprises from about 18.4% to about 27.6% by weight of totalphenolic antioxidants; e. from about 4.0% to about 6.0% by weight of thesupercritical extract of oregano, wherein the extract comprises fromabout 0.64% to about 0.96% by weight of total phenolic antioxidants; f.from about 10.0% to about 16.0% by weight of the hydroalcoholic extractof turmeric, wherein the extract comprises from about 5.6% to about 8.4%by weight of curcumin; g. from about 5.5% to about 8.0% by weight of thehydroalcoholic extract of rosemary, wherein the extract comprises fromabout 18.4% to about 27.6% by weight of total phenolic antioxidants; h.from about 10.0% to about 16.0% by weight of the hydroalcoholic extractof holy basil, wherein the extract comprises from about 1.6% to about2.4% by weight of ursolic acid; i. from about 10.0% to about 16.0% byweight of the hydroalcoholic extract of green tea, wherein the extractcomprises from about 36% to about 54% by weight of polyphenols; j. fromabout 8.0% to about 12.0% by weight of the hydroalcoholic extract ofhuzhang, wherein the extract comprises from about 6.4% to about 9.6% byweight of resveratrol; k. from about 4.0% to about 6.0% by weight of thehydroalcoholic extract of Chinese goldthread, wherein the extract fromabout 4.8% to about 7.2% by weight of berberine; l. from about 4.0% toabout 6.0% by weight of the hydroalcoholic extract of barberry, whereinthe extract from about 4.8% to about 7.2% by weight of berberine; and m.from about 2.0% to about 3.0% by weight of the hydroalcoholic extract ofScutellaria baicalensis; and wherein said composition further comprises:(i) the supercritical extract of ginger and the hydroalcoholic extractof ginger at a weight ratio of from about 0.9 to about 1.4 parts ofsupercritical extract per 1 part of post supercritical hydroalcoholicextract; (ii) the hydroalcoholic extract of turmeric and thesupercritical extract of turmeric at a weight ratio of from about 8 toabout 12 parts of hydroalcoholic extract per 1 part of supercriticalextract; and (iii) the supercritical extract of rosemary and thehydroalcoholic extract of rosemary at a weight ratio of from about 1.6to about 2.4 parts of supercritical extract per 1 part of hydroalcoholicextract.
 23. The method of claim 1, said composition is administered ina daily dosage of at least about 700 mg.
 24. The method of claim 1,wherein said composition is administered on a daily basis for at least 4weeks.
 25. The method of claim 5, wherein said at least one canceroustumor is detected during surgery on the central nervous system of saidsubject, having not been felt by a physician on physical examination ofsaid subject.
 26. The method of claim 5, wherein said at least onecancerous tumor is confined to the central nervous system of saidsubject and is detected by a physician on physical examination of saidsubject.
 27. The method of claim 5, wherein cancer related to said atleast one cancerous tumor extends beyond the central nervous system ofsaid subject, but has not spread to lymph nodes in said subject.
 28. Themethod of claim 5, wherein cancer related to said at least one canceroustumor is metastasized to regional lymph nodes or other parts of saidsubject.